Re: [Histonet] Causes of false positive Congo Red
Hi John, I must apologize again. We used to use the method from Carson's book. We now make up the reagents as follows: *Stock alkaline salt solution* Sodium chloride. 2g Distilled Water... 20mL Stir until the salt is dissolved, then with continuous stirring on a magnetic stirrer add 80mL of 100% denatured ethanol. Some salt may precipitate out after the ethanol is added. *Working alkaline salt solution * Stock alkaline stock solution...50 ml 1% Sodium Hydroxide0.5ml Filter and use within 15 minutes *Stock Congo red solution* Congo red 0.1g Stock alkaline salt solution 50mL Stir well with the magnetic stirrer and *let stand overnight or for a minimum of 3 hours* if the slides need to be ready the same day that the order was placed. *Working Congo red (Congo red)* Stock Congo red.. 50ml Sodium hydroxide 1%... 0.5ml Filter and use within 15 minutes. On Thu, Jun 20, 2024 at 9:31 AM Greg Dobbin wrote: > Good day John, > Very nice to hear from you again! I have been consulting your textbook in > my investigations! > Sorry about the brevity of the description of our method. I felt like my > post was already too long > and it might scare off some would-be contributors! :-) And yes, I > incorrectly referred to the dichroic green as "fluorescent"-thank you. > > Our method follows the Puchtler method described on pages 132-3 in Frieda > Carson's "Self-Instructional" textbook (1990) as does > the hospital that repeated our false-positive Congo Reds. Note, once we > re-made our reagents, our results returned to accurate staining. > Greg > > On Thu, Jun 20, 2024 at 2:49 AM John Kiernan wrote: > >> Greg, your method is incompletely described in your Histonet post, but it >> looks quite different from the "traditional" Highman's procedure (*Arch. >> Path*. *41*:559-562). What method were they using "at another >> lab" to get correct red amyloid that is green (dichroic, not fluorescent) >> with crossed polars? >> *John Kiernan* >> >> https://www.schulich.uwo.ca/anatomy//people/faculty/emeriti/kiernan_john.html >> = = = >> -- >> *From:* Greg Dobbin via Histonet >> *Sent:* June 19, 2024 8:53 AM >> *To:* histonet@lists.utsouthwestern.edu < >> histonet@lists.utsouthwestern.edu> >> *Subject:* [Histonet] Causes of false positive Congo Red >> >> Hello experts, >> *Some background:* >> I know that Congo Red can bind nonspecifically to non-amyloid components >> such as collagen and elastin under certain conditions (eg Carnoys >> fixative, >> insufficient differentiation, insufficient alkalinity, etc). However, >> everything I have been able to read on the topic suggests that >> over-staining is "easily" differentiated from true amyloid staining by >> using polarizing light microscopy. That is, true amyloid produces apple >> green fluorescence while non-amyloid components produce silver/grey color. >> >> *My question:* >> I want to know if anyone has encountered false positive staining that *is >> apple green* in color? We had a few bone marrow core biopsies that stained >> bright green but were later found to be negative when stained at another >> lab. We subsequently threw out all of our working solutions and made up >> everything fresh and repeated the previous (false positive) specimens and >> they were indeed negative in our lab as well. >> >> *In order to prevent this from happening again, I need to attempt to >> understand what may have caused this to happen in the first place. * >> >> This is where the vast collective knowledge of this group comes in. :-) >> Can anyone offer some insight as to possible causes? >> >> *Our Congo Red method:* >> >> >> Deparaffinize sections and bring them to water. >> >> Stain in Hematoxylin for 1 minute >> >> Add 0.5ml of 1% Sodium Hydroxide to 50 ml of stock alkaline salt solution. >> >> Wash slides in running water >> >> Place in *working* alkaline salt solution from step 2 for 20 minutes >> >> Add 0.5 ml of 1% Sodium Hydroxide to stock Congo red solution. >> >> Start to filter *working* Congo red solution when 15 mins are left in >> step 6 >> >> Place sections in the *working* Congo red from step #8 for 20 minutes. >> >> Dehydrate the slides one at a time in 3 changes of absolute ethanol, 6 >> dips >> each. >> >> Dip the slide 10 times in a coplin of xylene. >> >> Continue dehydrating the other slides. >> >> Coverslip the slides. >> >> *Greg Dobbin* >> 1205 Pleasant Grove Rd >> Route 220 >> York, PE C0A 1P0 >> ___ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Causes of false positive Congo Red
Good day John, Very nice to hear from you again! I have been consulting your textbook in my investigations! Sorry about the brevity of the description of our method. I felt like my post was already too long and it might scare off some would-be contributors! :-) And yes, I incorrectly referred to the dichroic green as "fluorescent"-thank you. Our method follows the Puchtler method described on pages 132-3 in Frieda Carson's "Self-Instructional" textbook (1990) as does the hospital that repeated our false-positive Congo Reds. Note, once we re-made our reagents, our results returned to accurate staining. Greg On Thu, Jun 20, 2024 at 2:49 AM John Kiernan wrote: > Greg, your method is incompletely described in your Histonet post, but it > looks quite different from the "traditional" Highman's procedure (*Arch. > Path*. *41*:559-562). What method were they using "at another > lab" to get correct red amyloid that is green (dichroic, not fluorescent) > with crossed polars? > *John Kiernan* > > https://www.schulich.uwo.ca/anatomy//people/faculty/emeriti/kiernan_john.html > = = = > -- > *From:* Greg Dobbin via Histonet > *Sent:* June 19, 2024 8:53 AM > *To:* histonet@lists.utsouthwestern.edu > > *Subject:* [Histonet] Causes of false positive Congo Red > > Hello experts, > *Some background:* > I know that Congo Red can bind nonspecifically to non-amyloid components > such as collagen and elastin under certain conditions (eg Carnoys fixative, > insufficient differentiation, insufficient alkalinity, etc). However, > everything I have been able to read on the topic suggests that > over-staining is "easily" differentiated from true amyloid staining by > using polarizing light microscopy. That is, true amyloid produces apple > green fluorescence while non-amyloid components produce silver/grey color. > > *My question:* > I want to know if anyone has encountered false positive staining that *is > apple green* in color? We had a few bone marrow core biopsies that stained > bright green but were later found to be negative when stained at another > lab. We subsequently threw out all of our working solutions and made up > everything fresh and repeated the previous (false positive) specimens and > they were indeed negative in our lab as well. > > *In order to prevent this from happening again, I need to attempt to > understand what may have caused this to happen in the first place. * > > This is where the vast collective knowledge of this group comes in. :-) > Can anyone offer some insight as to possible causes? > > *Our Congo Red method:* > > > Deparaffinize sections and bring them to water. > > Stain in Hematoxylin for 1 minute > > Add 0.5ml of 1% Sodium Hydroxide to 50 ml of stock alkaline salt solution. > > Wash slides in running water > > Place in *working* alkaline salt solution from step 2 for 20 minutes > > Add 0.5 ml of 1% Sodium Hydroxide to stock Congo red solution. > > Start to filter *working* Congo red solution when 15 mins are left in step > 6 > > Place sections in the *working* Congo red from step #8 for 20 minutes. > > Dehydrate the slides one at a time in 3 changes of absolute ethanol, 6 dips > each. > > Dip the slide 10 times in a coplin of xylene. > > Continue dehydrating the other slides. > > Coverslip the slides. > > *Greg Dobbin* > 1205 Pleasant Grove Rd > Route 220 > York, PE C0A 1P0 > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Causes of false positive Congo Red
Hello experts, *Some background:* I know that Congo Red can bind nonspecifically to non-amyloid components such as collagen and elastin under certain conditions (eg Carnoys fixative, insufficient differentiation, insufficient alkalinity, etc). However, everything I have been able to read on the topic suggests that over-staining is "easily" differentiated from true amyloid staining by using polarizing light microscopy. That is, true amyloid produces apple green fluorescence while non-amyloid components produce silver/grey color. *My question:* I want to know if anyone has encountered false positive staining that *is apple green* in color? We had a few bone marrow core biopsies that stained bright green but were later found to be negative when stained at another lab. We subsequently threw out all of our working solutions and made up everything fresh and repeated the previous (false positive) specimens and they were indeed negative in our lab as well. *In order to prevent this from happening again, I need to attempt to understand what may have caused this to happen in the first place. * This is where the vast collective knowledge of this group comes in. :-) Can anyone offer some insight as to possible causes? *Our Congo Red method:* Deparaffinize sections and bring them to water. Stain in Hematoxylin for 1 minute Add 0.5ml of 1% Sodium Hydroxide to 50 ml of stock alkaline salt solution. Wash slides in running water Place in *working* alkaline salt solution from step 2 for 20 minutes Add 0.5 ml of 1% Sodium Hydroxide to stock Congo red solution. Start to filter *working* Congo red solution when 15 mins are left in step 6 Place sections in the *working* Congo red from step #8 for 20 minutes. Dehydrate the slides one at a time in 3 changes of absolute ethanol, 6 dips each. Dip the slide 10 times in a coplin of xylene. Continue dehydrating the other slides. Coverslip the slides. *Greg Dobbin* 1205 Pleasant Grove Rd Route 220 York, PE C0A 1P0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC Validation Question
Hi Kara, Use multi-tissue block controls with normal tissues that are treated exactly the same (pre-analytically speaking) as your routine surgical specimens. Consult NordiQC.org for recommended normal control tissue for each IHC marker. The most used multi-tissue control block in my lab has pieces of tonsil, pancreas, small bowel and liver. *Greg Dobbin* 1205 Pleasant Grove Rd Route 220 York, PE C0A 1P0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Tisse disposal/formalin
Decant formalin off into formalin waste containers and the remaining tissue go into a red bag (red signifying destined for incineration). As the O.R. does with fresh tissues for disposal. *Greg Dobbin* 1205 Pleasant Grove Rd Route 220 York, PE C0A 1P0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] ISH Decal specimens
Use a formic acid based decal solution and MAKE SURE the specimen does not fix (formalin fixation) longer than 32hrs (false negatives result). Use 70% ETOH to manage the fixation time (ie fix for 24 hours, decal for 4 hrs, place in 70% ETOH until it gets put on the processor. Hold in 70% ETOH for weekends and holidays). Greg -- If God made a drink better than whiskey he kept it for himself! 🥃 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] destain for IHC
Hi Nancy, You should have no problem doing so. If you want proof, just run a couple of your IHC control sections through the H&E stainer as you would for a patient specimen (so right through to coverslipping) and then start the process of removing coverslip, hydration, destain in acid alcohol and then run slides on your IHC platform as you would -without the dewax step obviously. :-) *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *If God made a drink better than whiskey he kept it for himself! * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] cd138
We use the same. Our protocol is ER2 for 15 mins. We have 3 washes after polymer. Greg *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *If God made a drink better than whiskey he kept it for himself! * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Processing artefact??
Since images cannot be attached, if any of you feel you have some experience that I may find helpful...please email me directly and I will attach the image to my reply. Thanks again, *Greg Dobbin* Chief Technologist Anatomic Pathology Queen Elizabeth Hospital Charlottetown, PE, Canada ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Processing artefact??
Hello experts, Every now and then we will get stained slides that look like the attached image. We run one processing schedule (about 11 hrs) for everything so yes, our smaller biopsies are somewhat overprocessed, but 90-95% of the time they look perfectly fine. We can have GI biopsies in a case that go from A to P and find only one that looks like this. Moreover, recuts do not remedy the problem either...the problem seems to be occurring prior to microtomy. We use vendor grade Xylenes in our processor (a Leica ASP6025). All specimens are fixed for 18-24 hours prior to processing. Our GI blocks get a short soak in phenol prior to microtomy to help with the effects of overprocessing. All blocks within a case are treated the same. In fact we have also seen it in skin biopsies occasionally and skins do not get soaked in phenol. It is the randomness of the problem that is throwing us! Any ideas? Thanks, *Greg Dobbin* Chief Technologist Anatomic Pathology Queen Elizabeth Hospital Charlottetown, PE, Canada ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Tissue staining artefacts - areas of tissue not labelling
Hi Peter, We need more info: 1. What is the tissue(s)? Species? 2. What is the marker(s) in question? 3. Answer 1 and 2 with regard to the shared images? 4. What is the expected staining reaction? 5. Is there a control section placed on the same slide? If yes... 6. How does the control section perform? As expected or same artefact? *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *If God made a drink better than whiskey he kept it for himself! * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] ultrasafe formalin dispense
Honestly, I don't see the point of using this instrument in the histology lab (at least not my lab as we are set up). It would seem odd that we are being "ultra safe" when dispensing the formalin but then it's kind of the "wild west" when we have to discard the previously archived wet tissue! Obviously we are safe when discarding our used formalin but my point is that if there is no closed system for the discard process (none that I am aware of anyway) then why invest in the front end if the back end is still a mess! Now I can see Operating Rooms and Labour and Delivery suites loving this system as many nurses are less comfortable handling formalin. With this system they could safely fill their buckets and send the specimens off to us to let us worry about disposal. Just my 2 cents worth. Maybe someone on here will be able educate me about a safe system for tissue discards. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] formalin in OR
Hi Nancy, All routine specimens in our hospital are placed in formalin in the OR. Breast lumps and mastectomy specimens are sent up fresh so that they arrive STAT (to minimize cold ischemic times) and lymph nodes for lymphoma protocol are also sent up fresh. [*they actually send sentinel nodes up fresh too- it was just less confusing*] Diagnostic Imaging will place routine needle core biopsies directly in formalin (e.g needle cores of breast, prostate, non-lympoma lymph nodes and other misc. tissue masses). Lymph node cores for lymphoma protocol and renal cores are sent to the lab fresh, in a container on saline soaked Telfa pads. Endoscopy places all of their specimens directly in formalin. The derm clinic will place all routine skins directly into formalin but specimens destined for immunofluoresence are sent up to the lab fresh in a container on saline soaked Telfa pads. EBUS specimens are split between Cytology and Histology...the histo specimens go directly into formalin. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Fixation time and ISH
Background: I read on a vendor website that tissues stained using In situ hybridization should not exceed 32 hours fixation. Greater than 32 hours can produce false negative results because the validated retrieving protocol can be inadequate. We in fact have seen evidence of this in our lab (mystifying negative kappa/lambda staining) and we are beginning to realize that over-fixation May we’ll be the issue for us (eg weekend processing). I am proposing that we do what was common practice many years ago. That is, transfer the properly fixed and decalcified bone marrow cores into 70% ETOH until they can be processed into paraffin. My questions: Is anyone else doing this?? Is 70% ETOH still a viable option for labs in this situation?? And… Is there another idea and/or more information out there that could help us in this regard?? Thank you, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Cold ischemic times
This is more of a survey than a question: For those of you tracking and documenting your cold ischemic times for breast tissue (ie time out of body to time sliced [as needed] and immersed in formalin), and I assume most of you are...*what is your average time?* *Background:* *I ask because my director was looking for suggestions for quality indicators to report and while I feel like our cold ischemic average time is impressive at ~17 mins, she says that is pretty standard for everyone.* Thanks, Greg -- *Greg Dobbin* Chief Technologist - Anatomic Pathology Queen Elizabeth Hospital Charlottetown, PE Canada C1A 8T5 Ph: 902-894-2337 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] New cryostat validation procedure
Hello experts!
I would like to find out what others have done to validate a new cryostat.
I believe it would basically be the same as validating a new microtome
except specimens (ie blocks) are plentiful for microtomy whereas for us,
fresh tissue for cryosectioning is not readily available.
Background:
We are an acute care hospital and we only get about 50 intraoperative
consult requests per year. Our older unit is still functional so I can do
side-by-side comparisons with the new unit. Our frozens are only stained
for H&E and a very rare (one every couple of years) Oil Red O.
So I have two questions:
1. Can I use formalin-fixed tissue soaked overnight in a saturated
sucrose solution for the validation? This seems reasonable since I can do
side-by-side with the older unit. {autopsy tissue could be an option if the
timing coincided with our validation but Murphy's Law will probably prevent
this option! lol}
2. How many tissues would be satisfactory? Is one specimen of each of
our typical frozen specimen types enough? We typically get: ovary, thyroid,
skin and lymph node. Any other tissue type would be rare.
I look forward to hearing from you!
Thank you,
Greg
--
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE C0A 1P0
*Everything in moderation...even moderation itself**!*
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Re: [Histonet] Water under sections
Hi Bernice, In my lab, water under the sections is unique to charged slides. And you are correct, if there is water under the section when the slides are heated for antigen retrieval, the boiling (or at least very hot) water will damage or entirely destroy the section. We allow the charged slides to drain (upright) for a few minutes and then carefully make a hole in the edge of the wax and use a Kimwipe or paper towel to carefully wick the excess water out as much as we can without touching the tissue section. Then we "flick" or shake sharply to remove any residual water that may remain before we bake them (FYI, we choose not to bake on the stainer). If after baking 30 mins at 60C, we notice water (maybe someone was not as diligent earlier?), we wick away the excess and bake for another 15 mins to ensure good adhesion of the section to the slide before proceeding to the immunostainer. Hope this helps. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC validation after a new tissue processer is installed
Hi martha, Prognostic markers must be re-validated (Eg.s Breast markers and CD117) as you described. Every Ab in your menu should be tested (as you would for a new a new lot) and do not forget to validate your H&E (with various tissue types) and SS as well. For the H&Es, if possible do side-by-side comparisons between old and new processors (the downside of what is otherwise an exciting time!). Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Blank Spots on IHC- Bubbles on Bond
Hi Tim, The usual source of bubbles is a deteriorating syringe or lines or valve. Have you observed the syringe while running the Clean Fluidics maintenance function? There can be some "micro" bubbles in the syringe on first draw of a reagent however, if working properly these tiny bubbles quickly disappear in subsequent draws of that reagent. If bubbles persist in the barrel of the syringe throughout the flushing, then you have found the source of the problem. If the bubbles occur with every reagent then the syringe is the problem but if you only see bubbles in one of the reagents then it is a defective valve or line for that reagent. I hope this was helpful. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] process formalin-fixed tissues from animals infected with a virus
Very interesting paper John! Thank you. I wish the authors had also experimented with higher concentrations of formaldehyde (eg 10% formalin). Might one infer that 10% would be even more efficient in inactivating viral infectivity than 2 and 4%? 🤔 Cheers Greg On Thu, Aug 6, 2020 at 11:02 AM John Garratt wrote: > Evaluation of Virus Inactivation by Formaldehyde to Enhance Biosafety of > Diagnostic Electron Microscopy > > > https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4353909/ > > > It is nice to have a reference. > > > > John > > On Thu, Aug 6, 2020 at 4:10 AM, Greg Dobbin via Histonet < > histonet@lists.utsouthwestern.edu> wrote: > > Hi Amy, > Formalin fixed tissue is no longer infectious...unless you are talking > about prions (eg scrapie, BSE, etc). So there should otherwise be no > concerns or additional precautions required. > Cheers, > Greg > > -- > *Greg Dobbin* > 1205 Pleasant Grove Rd > <https://www.google.com/maps/search/1205+Pleasant+Grove+Rd?entry=gmail&source=g> > RR#2 York, > PE C0A 1P0 > > > *Everything in moderation...even moderation itself**!* > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] process formalin-fixed tissues from animals infected with a virus
Hi Amy, Formalin fixed tissue is no longer infectious...unless you are talking about prions (eg scrapie, BSE, etc). So there should otherwise be no concerns or additional precautions required. Cheers, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Spectra Staining Kits
Hi Wanda, If you have just purchased the Spectra stainer recently (ie still under the first year warranty), demand that Leica send their Application Specialist back on site. if they promised 2000 slides with no drop in quality over that period (I believe it is 2000 slides or 7 days, whichever come first)...then hold them to it! Make them work for you. You paid a lot of money for that system. You should not need to be on here looking for help. I have a Spectra and if you want to chat with me about our experience feel free to call me. I'll send you my number in a separate email. All the best, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Tissue Tek embedding centers
Hi Charles, If memory serves, there is a thermal fuse or switch that will shut the unit down if it exceeds a certain temp. Once the unit cools, the switch is reset. So either that switch needs to be replaced or there is a problem elsewhere that is causing the unit to overheat. Replacing that switch should be an easy and cost-effecient thing to try first. Good luck, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC troubleshooting
Nancy, You used the word “blot” in your reply to Charles, and while I can’t be certain that you did not mean the same...I would like say in the interest of clarity, that it is safer to carefully “wick” the water away from under each section prior to baking. Blotting (like we might do after a Gram stain) would likely remove parts or all of the section. All the best, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC Troubleshooting
Hi Charles, It adhesion problems can arise from several sources, some have already been mentioned. Here are two that I have experienced: 1) Bad slides. Either a manufacturing defect such that the positive charge is insufficient or the slides were somehow compromised after opening them on the bench (perhaps exposed to moisture thus removing the charge). Sometimes a new lot number be better but sometimes switching brands is the quickest fix. 2) Ensuring all water is removed from under the section. I find that with some charged slides (many in fact), water is trapped under the section and does not drain out on standing and does not drain or evaporate when baking at 60C. If the slide with trapped water is put on the instrument with trapped water, the dewax step will lift much of the tissue. Cheers, -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Thyroglobulin TGB04+TGB05 Not Working in Canine and Feline
Hi Sandra, >From your post, it looks to me like you are assuming the recommended dilution should work. I apologize in advance if I have misinterpreted your post in this regard. When starting with a new antibody (including a new vendor for same Ab and/or a new clone), do a range of dilutions that includes the recommended one. So in this case, I would try 1:50, 1:100 and 1:200 to start. Use the recommended AR first and then see how each of the 3 dilutions look. Just as an example, you may find 1:50 has a weak signal and higher dilutions are negative. If so, set up 3 more dilutions with the same AR (eg 1:10, 1:25 and 1:50). Once you get close to where you think the intensity should be you can try modifying the AR protocol to improve the sensitivity. Good luck, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Processing Schedule- ASP-6025
Great point John! We do attempt to have all of the specimen handling for our control tissue, match that of our specimens. However, tonsillectomies at our hospital are only performed on Thursdays. So we fix them for 24hrs (until Friday) then process overnight and remove from the processor on Saturday (we don't typically work Saturdays). And I have done this a couple of times but can I swear that this particular tonsil was handled that way? I cannot ...which I know, points to another deficiency! :-0 It could very well be that the fixation times of the two tonsils I am comparing are sufficiently different that they will require different retrieval times to produce stains of similar intensity. I also appreciate getting your opinion on the processing schedule. You have much more experience than I!! Thank you for your insights. All the best, Greg On Wed, May 8, 2019 at 10:50 AM John Garratt wrote: > The processing schedule looks fine. I am thinking that if you are having > no problems with morphology on the H&E processing is not the issue. The > extended period in the warming draw is a good hypothesis. > I do have a question. How long was that particular piece of tonsil kept in > formalin before processing? Could over fixation be the issue? > I have found that maintaining tight control of control tissues is > important which includes minimum and maximum fixation time. > > John > > Sent from ProtonMail Mobile > > > On Tue, May 7, 2019 at 5:52 PM, Greg Dobbin via Histonet < > histonet@lists.utsouthwestern.edu> wrote: > > Fascinating Tony! > We don’t typically leave them in the warming drawer any longer than a > couple of hours, but maybe this particular tonsil was left linger for some > reason and no one thought anything of it?! Something to consider for sure! > Thanks. > Greg > > On Tue, May 7, 2019 at 9:37 PM Tony Henwood (SCHN) < > tony.henw...@health.nsw.gov.au> wrote: > > > Processing seems adequate. > > > > After processing, how long do they sit in the embedding centre block > > holding tank before embedding? > > > > We found that quite a few antigens were affected when we stored control > > tonsil in the embedding centre (dry) at 60oC for a few days before > > embedding. In summary: > > > > Antibody Clone Dried (Normal = 3+) > > CD4 4B12 0 > > BOB-1 TG14 0 > > CD3 LN10 1+ > > CD79a JCB117 1+ > > Oct-2 Oct-207 1+ > > CD8 4B11 2+ > > CD20 L26 3+ > > > > So CD20 was unaffected but this process affected most of the antigens > with > > some losing antigen recognition by the antibody (eg CD4 and BOB-1). > > > > Another one of those pre-analytical issues we need to consider. > > > > And yes I am writing this up for submission! > > > > > > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) | > > Principal Scientist; Adjunct Fellow, School of Medicine, University of > > Western Sydney; Visiting Lecturer, School of Life Sciences, Faculty of > > Science, University of Technology Sydney | Histopathology > > t: (02) 9845 3306 | f: (02) 9845 3318 | e: > tony.henw...@health.nsw.gov.au > > | w: www.schn.health.nsw.gov.au > > m: > > > > > > Cnr Hawkesbury Road and Hainsworth Street, Westmead, NSW Australia > > Locked Bag 4001, Westmead 2145, NSW Australia > > > > ♲ Please consider the environment before printing this email. > > > > -Original Message- > > From: Greg Dobbin via Histonet [mailto:histonet@lists.utsouthwestern.edu > ] > > Sent: Wednesday, 8 May 2019 5:07 AM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] Processing Schedule- ASP-6025 > > > > Hello colleagues, > > I recently stained (IHC) a section of normal tonsil from another facility > > with p16 and the resulting stain was better than the same stain on a > > section of my labs own normal tonsil control. > > > > This has led us to question our processing schedule. I am not concerned > > with our fixation because we fix everything for at least 24 hours in 10% > > formalin (commercially prepared) prior to processing. > > > > Does anything jump out at you as being a potential red flag in the > > following overnight protocol? > > > > - Formalin 15 mins; RT > > - Processing water 1 min; RT > > - ETOH 70% 30 mins; 35C > > - ETOH 80% 30 mins; 35C > > - ETOH 95% 30 mins; 35C > > - ETOH 100% 30 mins; 35C > > - ETOH 100% 40 mins; 35C > > - ETOH 100% 60 mins; 35C > > - Xylene 60 mins; 35C > > - Xylene 60 mins; 35C > > - Xylene 60 mins; 35C > > - Paraffin 60 mins; 57C; vacuum > > - Paraffin 60 m
Re: [Histonet] Processing Schedule- ASP-6025
Fascinating Tony! We don’t typically leave them in the warming drawer any longer than a couple of hours, but maybe this particular tonsil was left linger for some reason and no one thought anything of it?! Something to consider for sure! Thanks. Greg On Tue, May 7, 2019 at 9:37 PM Tony Henwood (SCHN) < tony.henw...@health.nsw.gov.au> wrote: > Processing seems adequate. > > After processing, how long do they sit in the embedding centre block > holding tank before embedding? > > We found that quite a few antigens were affected when we stored control > tonsil in the embedding centre (dry) at 60oC for a few days before > embedding. In summary: > > AntibodyClone Dried (Normal = 3+) > CD4 4B120 > BOB-1 TG140 > CD3 LN101+ > CD79a JCB117 1+ > Oct-2 Oct-207 1+ > CD8 4B112+ > CD20L26 3+ > > So CD20 was unaffected but this process affected most of the antigens with > some losing antigen recognition by the antibody (eg CD4 and BOB-1). > > Another one of those pre-analytical issues we need to consider. > > And yes I am writing this up for submission! > > > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) | > Principal Scientist; Adjunct Fellow, School of Medicine, University of > Western Sydney; Visiting Lecturer, School of Life Sciences, Faculty of > Science, University of Technology Sydney | Histopathology > t: (02) 9845 3306 | f: (02) 9845 3318 | e: tony.henw...@health.nsw.gov.au > | w: www.schn.health.nsw.gov.au > m: > > > Cnr Hawkesbury Road and Hainsworth Street, Westmead, NSW Australia > Locked Bag 4001, Westmead 2145, NSW Australia > > ♲ Please consider the environment before printing this email. > > -Original Message- > From: Greg Dobbin via Histonet [mailto:histonet@lists.utsouthwestern.edu] > Sent: Wednesday, 8 May 2019 5:07 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Processing Schedule- ASP-6025 > > Hello colleagues, > I recently stained (IHC) a section of normal tonsil from another facility > with p16 and the resulting stain was better than the same stain on a > section of my labs own normal tonsil control. > > This has led us to question our processing schedule. I am not concerned > with our fixation because we fix everything for at least 24 hours in 10% > formalin (commercially prepared) prior to processing. > > Does anything jump out at you as being a potential red flag in the > following overnight protocol? > >- Formalin 15 mins; RT >- Processing water 1 min; RT >- ETOH 70% 30 mins; 35C >- ETOH 80% 30 mins; 35C >- ETOH 95% 30 mins; 35C >- ETOH 100% 30 mins; 35C >- ETOH 100% 40 mins; 35C >- ETOH 100% 60 mins; 35C >- Xylene 60 mins; 35C >- Xylene 60 mins; 35C >- Xylene 60 mins; 35C >- Paraffin 60 mins; 57C; vacuum >- Paraffin 60 mins; 57C; vacuum >- Paraffin 60 mins; 57C; vacuum > > Our formalin is changed after every 1100 cassettes and the alcohol, > xylenes and paraffins are managed similarly by the instrument. Our specimen > mix is a little of everything (skins, GIs, breasts, needle cores, gall > bladders, gyne, etc). > > The one unknown (so far) in this story, is how the tonsil from the other > laboratory was handled (ie the fixative used and for how long-I am assuming > 10% formalin). > > Obviously, many of you will have schedules that differ from this one, in > any number of ways, but what I am looking for from you is your opinion: *is > there anything about this schedule that is particularly concerning?* Thank > you, Greg > > > -- > *Greg Dobbin* > 1205 Pleasant Grove Rd > <https://maps.google.com/?q=1205+Pleasant+Grove+Rd&entry=gmail&source=g> > RR#2 York, > PE C0A 1P0 > > > *Everything in moderation...even moderation itself**!* > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > This message is intended for the addressee named and may contain > confidential information. If you are not the intended recipient, please > delete it and notify the sender. > > Views expressed in this message are those of the individual sender, and > are not necessarily the views of NSW Health or any of its entities. > -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Processing Schedule- ASP-6025
Hello colleagues, I recently stained (IHC) a section of normal tonsil from another facility with p16 and the resulting stain was better than the same stain on a section of my labs own normal tonsil control. This has led us to question our processing schedule. I am not concerned with our fixation because we fix everything for at least 24 hours in 10% formalin (commercially prepared) prior to processing. Does anything jump out at you as being a potential red flag in the following overnight protocol? - Formalin 15 mins; RT - Processing water 1 min; RT - ETOH 70% 30 mins; 35C - ETOH 80% 30 mins; 35C - ETOH 95% 30 mins; 35C - ETOH 100% 30 mins; 35C - ETOH 100% 40 mins; 35C - ETOH 100% 60 mins; 35C - Xylene 60 mins; 35C - Xylene 60 mins; 35C - Xylene 60 mins; 35C - Paraffin 60 mins; 57C; vacuum - Paraffin 60 mins; 57C; vacuum - Paraffin 60 mins; 57C; vacuum Our formalin is changed after every 1100 cassettes and the alcohol, xylenes and paraffins are managed similarly by the instrument. Our specimen mix is a little of everything (skins, GIs, breasts, needle cores, gall bladders, gyne, etc). The one unknown (so far) in this story, is how the tonsil from the other laboratory was handled (ie the fixative used and for how long-I am assuming 10% formalin). Obviously, many of you will have schedules that differ from this one, in any number of ways, but what I am looking for from you is your opinion: *is there anything about this schedule that is particularly concerning?* Thank you, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Histonet Digest, Vol 184, Issue 23
Hello all, While I do not disagree with anything that has been said thus far regarding the failed refrigerator and moving forward, I think Tim Morken's information and shared experience was the most relevant! I know that my Bond instruments are running 24 hrs a day, so some of the more commonly ordered antibodies and certainly our detection kits (enzyme included) can be out of the fridge for days with no noticeable deterioration. :-) Greg > > -- Forwarded message -- > From: MARY ANN > To: > Cc: > Bcc: > Date: > Subject: [Histonet] Fridge temp > Let's say, hypotheticaly, if you discover your fridge with all you > antibodies and detection kits were discovered to have been at 19c. for an > undisclosed amout of time. 12 24 48 hours due tona power surge..south > Florida weather. > > Let's also propose your lab CFO/Owner dosent think its a big deal. > > First how would you handle the issue given the frisge has been restored ? > > > > -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] ER/PR question
Hi Karen, As mentioned by others "decay" is not likely going to be an issue. More concerning for you could be not knowing how those tissues were handled prior to processing 10 years ago. Presumably, you now track cold ischemic times and have standardized your fixation protocols for breast tissues and you have validated your IHC procedures with these current parameters. If your lab was like many others 10 years ago, these parameters were probably not maintained so rigorously. So interpretation of a negative result may be problematic. Having said that, there is the internal control for both ER and PR. It seems to me that if the internal control is staining adequately, then perhaps interpretation will not be an issue. I am curious, does anyone have a different take on relying the internal control as a guide in this particular situation?? Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC-H&E-IHC HELP...
Hi Curt, You can de-stain the hematoxylin counter stain and then do H&E and all should be fine. Subsequent IHC is possible but obviously you would need a different color chromogen to differentiate the new stain from the previous one. There may be a problem getting the section stay on through a second IHC procedure but maybe not. I would stain a control section for the desired Ab using the retrieval method that was used initially and see how the control looks. If the pathologist thinks the that this control is adequate for interpretation, then restain the slides with the desired Ab and no additional retrieval. Cheers, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Question - EM
Hi Dr. Cartun, It has been many years since I worked in EM but I my recollection is that tissues could remain in 2% Glut indefinitely without detriment (for EM purposes). However, Osmium tetroxide had to have a limited exposure. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Leica Bond Max artifact
Hi Linda, Is this a problem that has just started or has the red kit always had this artefact? - If it just started, perhaps it is a problem with a particular kit lot number (or numbers). If so, get Leica Tech service involved to see if anyone else has reported a problem. - If it has been an ongoing issue for the red kit then perhaps you need to modify the Red Kit protocol by adding additional (and/or longer) wash steps. Here again Tech Service can help you with this. - The only other thing I can think to ask is: Do you have anyone new operating the instrument? Could a new person be putting undiluted Bond Wash in the bulk container? Let us know what you find out. Good luck. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Braf protocol on Bond-III
1:100 with ER-2 for 30 mins. Clone is the V600E from Spring BioScience Looks great for uis and we are performing well on EQA surveys. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] BRAF
Yes, it works. Very well actually. And we have done well on both a cIQc and a CAP survey. Greg On Tue, Jul 24, 2018 at 3:20 PM Alminawi, Samira < samira.almin...@sunnybrook.ca> wrote: > Hi Greg, > > How is the quality of the staining? Is it working for melanoma? > > Samira > > > > *From:* Greg Dobbin [mailto:greg.dob...@gmail.com] > *Sent:* Tuesday, July 24, 2018 2:17 PM > *To:* Alminawi, Samira; histonet@lists.utsouthwestern.edu > *Subject:* RE: BRAF > > > > Here in Charlottetown we are using V600E (VE-1) clone (Monoclonal) from > Spring Diagnostics. We run it on the Bond platforms at 1:100 with ER-2 > (high pH) for 30 mins. > > Greg > > > > -- > > *Greg Dobbin* > 1205 Pleasant Grove Rd > RR#2 York, > PE C0A 1P0 > > > > *Everything in moderation...even moderation itself!* > > *This e-mail is intended only for the named recipient(s) and may contain > confidential, personal and/or health information (information which may be > subject to legal restrictions on use, retention and/or disclosure). No > waiver of confidence is intended by virtue of communication via the > internet. Any review or distribution by anyone other than the person(s) > for whom it was originally intended is strictly prohibited. If you have > received this e-mail in error, please contact the sender and destroy all > copies.* > > > -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] BRAF
Here in Charlottetown we are using V600E (VE-1) clone (Monoclonal) from Spring Diagnostics. We run it on the Bond platforms at 1:100 with ER-2 (high pH) for 30 mins. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] USA to Canada
Pass The General MLT Certification exam, as our MLT exam includes Histology and Microbiology. :-) Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Slide and block retention -Peds seperated?
Hello all, I would like to find out what other clinical path labs are doing in Canada in particular but in the US as well with regard to retention of blocks and slides. Are other labs filing pediatric cases separately from adult cases? I am required to archive specimens from pediatric cases separately from adults because our retention schedule states that specimens for adults will be retained for *20 years* while specimens from pediatric cases will be retained for *20 years after the patient reaches the age of majority* (18 years old). So for example, a specimen on a newborn will be kept for 38 years while a specimen on a 17-year-old will be kept for 21 years. This is no small task to achieve. We must generate a list of pediatric cases 4 times a year, pull these cases out of the file, file them by birth year in another area separate from our regular archived material, maintain and update a separate hardcopy case list, etc. I am proposing to management that we keep all cases for 38 years so that we can eliminate this task of separating peds from adults. However, before my request will be entertained, I must be able to point to other path labs that are not separating their peds cases. Thank you for your time! Greg -- *Greg Dobbin, MLT* Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Leica open containers
Hi Charles, If you are using the Leica Diluting Buffer or another commercially prepared diluting buffer the antibody should remain stable for a very long time. I can't say how long, I suspect there would be numerous variables to be considered but generally...I would expect very little loss of intensity over two years even. For me, the antibody expires long before the any deterioration of signal is noticed. In your case, if you are using a commercially prepared diluent, then start looking at extended times left out of the fridge, a malfunctioning fridge, contamination, etc. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] p16 Antibody
Hi Laurie, We are purchasing the antibody only from Roche/Ventana and optimizing it for use with our detection system (Bond Refine Detection kit) and the Bond-III stainer. Ask your Ventana person about buying the Ab only. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Paraffin infiltration issue, will this impact IHC?
Hi Merissa, As far as I am aware, insufficient paraffin infiltration would only affect sectioning. The epitopes that we we are attempting to stain with IHC are affected by pre-analytic factors such as fixation, cold ischemic time and perhaps heat (too much) but plus or minus wax should not be an issue if you are able to obtain good sections. Have you stained these poorly infiltrated specimens with H&E? Are you certain you have good sections with all tissue components represented? That would be my main concern. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Leica embedding station
Hi Lauren, Further to Tony's response...there are also thermal fuses that shut the power off to a drawer if the power (or temp IDK which) as a safety feature. So quite often we find one warming drawer or the other can be reset by powering the unit down for a couple of hours. It normally works fine after that...but not always. When powering down doesn't fix it we call biomed engineers to come with their volt meters and spare fuses to fix it. We have also had heating elements "die" and in that case the whole drawer must be replaced because the element is affixed to the bottom of the drawer. Hope this helps, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Antibody Validation CLIA
Hi Paula, Let me first say I am Canadian and my lab is not governed by CLIA or CAP. But for what it is worth, here are my thoughts... When thinking about validating antibodies for IHC you must first consider whether the antibody in question is Class-I (prognostic eg Breast markers, CD117, etc) or Class-II. Class-I require a more robust validation protocol then Class-II antibodies. Next you need to consider why the validation is needed. Is it: 1. a new Ab in your lab? 2. a new vendor for an antibody? 3. a new lot number of an existing antibody in your menu? 4. a change in protocol of an existing antibody? #'s 1 and 2 require a larger validation whereas #'s 3 and 4 should only require a minimal check (unless of course there is a drastic change in the protocol in #4). As for absolute numbers of slides for each, unless CLIA provides this information, you are left to work with your Director to find that balance between cost (ie more slides = increased cost) with confidence (how many do you need to see in order to feel confident that the stain is performing as expected). I hope this helps. Sincerely, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] re; cell blocks and frozen for IHC
No pretreatments for anything that is not formalin fixed. I think 95% ETOH for a fixative but but others may have a better idea than I. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] AFB Stain on Cytospin Preps
Good day colleagues, We are occasionally being asked to do an acid-fast stain (Ziehl-Neelsen) on a alcohol-fixed cytospin preparation from a cytology fluid. Typically in Cytology, if a specimen is very bloody, we will add acetic acid to lyse the red cells. Does anyone know if acetic acid would have any adverse affect on the stainability of AFB organisms if present? Thank you, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Kappa and Lambda by ISH
Hi Folks, As it happens, we had a lymphoma case this week that was Kappa restricted (confirmed at a consulting laboratory by Flow Cytometry) and our Kappa and Lambda ISH staining was negative for both! In discussing it further with our Hematopathologist, our ISH probes for Kappa and Lambda in bone marrow specimens only stains plasma cells it does *NOT* stain B-cells. So this is likely why LN tend to be negative. Maybe one of our Histonet-contributing pathologists would like to elaborate? :-) Greg -- *Greg Dobbin* Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Kappa & Lambda ISH
Hello Renee, We do K & L by ISH on the Bond-III and BondMax with Refine detection without issue on any of the tissues we have run (GI, skin, BM, LN). We have a normal tonsil control section on every slide. Do you also run a positive control on every slide? How does it perform? Next step would be for me to share our protocol. Let me know if you would like me to send that to you. Cheers, Greg -- *Greg Dobbin, R.T.* Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Storage of blanks (Charged slides)
Hi Folks, Can some of you share what your practices are regarding storage of blanks for possible subsequent IHC orders? Background: We cut a blank on every prostate core in case a PIN4 is ordered after the H&Es have been read. So each week, we have 72-100 blank slides to store. Space is an issue especially if we keep them in a freezer. Due to space constraints, we are currently storing these blank slides in slide filing fashion (ie front-to-back) in the -20 freezer. Questions: 1) How long are you keeping your blank slides? 2) Under what storage conditions are you keeping then (ie Temp, in a box, wrapped, stacked, etc)? Thank you, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Section adherence issues (IHC)
Hi Folks, I haven't been on here much lately but it is nice to know that we are all here for each other when needed! My problem is section adherence during IHC staining. And it is not all of the time it is intermittent; so not all of the time and not on all slides when it does occur. Background: We have a Bond-III immunostainer and we use the Leica Apex charged slides for our IHC stains. We have no additives in our water baths. Our sections drain in a stand and then any trapped water under the section is either flicked or wicked away as needed prior to placing the slides in a rack in a 60 C oven for 30 mins prior to staining. We do not do on board baking (primarily because on board baking is only 10 mins long and with the section lifting issue we want longer). Some of the specimen types are more susceptible it would seem. Cervix LEEP specimens tend to be bad, we use a 4mm punch to obtain some of our control tissue and so the breast tissue we use for Myosin Heavy Chain seems to be a bad one and sometimes our ER/PR control sections (but not always). Fine needle cores where a lot of tumour is present tend to be bad (again not always). We have tried baking longer, we turn ourselves inside out trying to get all of the water out from under the sections, we have tried charged slides from another manufacturer. I have looked at the HIER protocols and none are extraordinary in nature. We use very clean covertiles (no scratches or blemishes). Our specimens are fixed for 24hrs before processing. We use the same slides for Special Stains and don't have this issue there. I need some new suggestions to try! All ideas welcome. Cheers, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Releasing of Patient Tissue
Vanessa, We strongly encourage people that make such requests to use a funeral home or crematorium to handle the tissue for them (for example an aborted fetus; or some cultures wish to have an amputated limb buried in their future burial plot). If they take their own amputated limb and then were to carelessly or accidentally discard it, a whole host of resources will be wasted investigating the origin of the found tissue! If they insist, we have a waiver that they must sign and then we rinse the tissue very well with running water to remove most of the residual formalin before packing it up for pick up. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] grad student problem
I can't speak to the the T-cell receptor sites but the only option for getting decent sections at this point is option number two. In the future, (if there applicable) the cut surface of a cryosectioned block can be recovered with OCT, frozen, and stored in an air-tight whirl-pak or ziplock bag in the freezer for a little longer to avoid freezer burn. Cheers, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Uneven ER/PR
Joanne, I do not think you should expect to get perfectly uniform IHC staining in breast core biopsies every time. At our hospital, we prefer to avoid doing hormone receptor stains on breast cores because they are not as "robust" as a lumpectomy or mastectomy specimen. The unevenness could be due to pre-analytic causes beyond your control (such as: are they placed in directly and immediately into formalin by the radiologist? Is your processing schedule optimized for small biopsies or is it a "one-size fits all" schedule for large and small specimens, etc). Occasionally, we will agree to an Oncology request to stain them but this is usually only done when they feel it is not going to be clinically beneficial to put the patient through a more invasive procedure (ie lumpectomy). Another disadvantage of looking at cores vs lump specimens is that you cannot see the "bigger picture". Perhaps there is heterogeneity in the intensity of the ER/PR expression throughout the tumor; something that is easier to discern when you can see the staining pattern across the entire cross-section of the tumor. I too use the Bond platform. If you are placing appropriate tissue controls on the same slide as the patient and these have the expected staining pattern and intensity, you may be fairly certain that the issue was (or is) pre-analytic in nature. If this is not your practice, then try re-staining new sections of the same case. If it is corrected, contact Leica Tech support to try to figure out what went wrong the first time. If it is not corrected, then you are back to suspecting a pre-analytic issue (assuming you are not seeing this uneven-ness in other IHC specimens). I strongly encourage use of tissue controls on every patient slide. Cheers, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC Staining (Pardue, Judith)
Judith, The red chromagen is suseptible to "fading" if stained slides are left in either alcohol or xylene for too long. Check to make sure that everyone involved is following the same procedure for the dehydraytion, clearing and mounting of the stained slides. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 med*lab*professionals.ca *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Safety question
Buy yourself a roll of WHMIS labels. Get ones that are sufficiently large to be easily read while still fitting on the side of the secondary container. All necessary info will be captured there once you check the right boxes, etc. Cheers, Greg -- Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Microtomy Statistics
Are you hoping to measure competency or productivity? If it is competency that you hope to assess, allow your tech to cut blocks of a variety of specimen types for 1 hour *without* distractions. If they cut 30 or more blocks and produce good quality sections, then they are meeting your standard. If it is productivity that you want to assess, then you may want to look at an average of several daily totals per tech. If you need to establish what the desired minimal productivity benchmark is for your lab, do this for several or all techs and come up with a reasonable (and achievable) benchmark for all to strive for as their minimum. Cheers, Greg -- Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Looking for fine mesh cassette
Hi Folks, I am having difficulty finding a fine mesh biopsy cassette that is compatible with my TBS cassette printer (i.e. *45 degree angle* print surface). The holes in the TBS cassette that I am currently using are too big (~1mm). Can anyone suggest a supplier with a good quality cassette that will work with my printer?? Thank you, Greg Dobbin Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
histonet@lists.utsouthwestern.edu
Are you soaking your blocks in dilute ammonium hydroxide before sectioning? Soaking too long will affect stain quality. Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "Jordan, Velma" 03/15/11 22:22 PM >>> We have been troubleshooting complaints of hematoxylin staining light in our H&E. The main complaint is no differentiation between the nucleus and cytoplasm. We are using Richard-Allan hematoxylin ,eosin,clarifier and bluing. We tried surgipath, changing times, pH the H2O and any other thing we could think of, but the problem remains. Does anyone have a suggestion? Velma Jordan UHS Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient (s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution of this information is prohibited. If this was sent to you in error, please notify the sender by reply e-mail and destroy all copies of the original message.___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] negative controls on immunos
Thanks Joyce. This excerpt supports what I had said- noting the difference between the negative (or deletion) control (which is what I was addressing in my reply) and the negative tissue control. Cheers! Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "Weems, Joyce" 3/2/2011 11:30 AM >>> The CAP guidelines are pretty clear. Copied from latest checklist.. Isn't this fun???! j:>) ANP.22570 Phase IIN/A YES NO Are appropriate negative controls used? NOTE: Negative controls must assess the presence of nonspecific staining in patient tissue as well as the specificity of each antibody. Results of controls must be documented, either in internal laboratory records, or in the patient report. A statement in the report such as, "All controls show appropriate reactivity" is sufficient. A negative reagent control is used to assess nonspecific or aberrant staining in patient tissue related to the antigen retrieval conditions and/or detection system used. A separate section of patient tissue is processed using the same reagent and epitope retrieval protocol as the patient test slide, except that the primary antibody is omitted, and replaced by any one of the following: ■ An unrelated antibody of the same isotype as the primary antibody (for monoclonal primary antibodies) ■ An unrelated antibody from the same animal species as the primary antibody (for polyclonal primary antibodies) ■ The negative control reagent included in the staining kit ■ The diluent/buffer solution in which the primary antibody is diluted In general, a separate negative reagent control should be run for each block of patient tissue being immunostained; however, for cases in which there is simultaneous staining of multiple blocks from the same specimen with the same antibody (e.g., cytokeratin staining of multiple axillary sentinel lymph nodes), performing a single negative control on one of the blocks may be sufficient provided that all such blocks are fixed and processed identically. This exception does not apply to stains on different types of tissues or those using different antigen retrieval protocols or antibody detection systems. The laboratory director must determine which cases will have only one negative reagent control, and this must be specified in the department's procedure manual. The negative reagent control would ideally control for each reagent protocol and antibody retrieval condition; however, large antibody panels often employ multiple antigen retrieval procedures. In such cases, a reasonable minimum control would be to perform the negative reagent control using the most aggressive retrieval procedure in the particular antibody panel. Aggressiveness of antigen retrieval (in decreasing order) is as follows: pressure cooker; enzyme digestion, boiling; microwave; steamer; water bath. High pH retrieval should be considered more aggressive than comparable retrieval in citrate buffer at pH 6.0. It is also important to assess the specificity of each antibody by a negative tissue control, which must show no staining of tissues known to lack the antigen. The negative tissue control is processed using the same fixation, epitope retrieval and immunostaining protocols as the patient tissue. Unexpected positive staining of such tissues indicates that the test has lost specificity, perhaps because of improper antibody concentration or excessive antigen retrieval. Intrinsic properties of the test tissue may also be the cause of "non-specific" staining. For example, tissues with high endogenous biotin activity such as liver or renal tubules may simulate positive staining when using a detection method based on biotin labeling. A negative tissue control must be processed for each antibody in a given run. Any of the following can serve as a negative tissue control: 1. Multitissue blocks. These can provide simultaneous positive and negative tissue controls, and are considered "best practice" (see below). 2. The positive control slide or patient test slides, if these slides contain tissue elements that should not react with the antibody. 3. A separate negative tissue control slide. The type of negative tissue control used (i.e., separate sections, internal controls or multitissue blocks) should be specified in the laboratory manual (refer to ANP.22250). Multitissue blocks may be considered best practice and can have a major role in maintaining quality. When used as a combined positive and negative tissue control as mentioned above, they can serve as a permanent record documenting the sensitivity and specificity of every st
Re: [Histonet] negative controls on immunos
Yes, it is always best to run every conceivable control available. Then you can be really, really, really sure. However, if you have practical issues that come into play such as cost of reagents, or extra controls taking up valuable space on the stainer then you might have to think about what you are trying to accomplish with each control and balance the cost vs benefit. So what are we checking with the negative control? We are checking the reagents to ensure that there is no non-specific reaction arising from the detection kit or the buffers, etc. If you are running the same block tomorrow with the same detection kit (ie same lot) it is not necessary (in my humble opinion) to check it again with another negative control. I suppose if you are worried that someone could be trying to sabotage your work by sneaking into your lab at night and contaminating your detection kit...then the simplest way to detect this sinister activity would be to run a negative control every time. Otherwise, I think one negative control slide per block per detection kit will be adequate. Disclaimer: Please know that I have just tried to inject a little humor into this response. I am really not trying to be sarcastic in anyway. Cheers! Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "Diana McCaig" 3/1/2011 2:07 PM >>> If you do a run of several immunos today and you run a negative control, and there is a request for additional immunos tomorrow, would you run another negative control with the additional slides. They are being stained on a stainer and not manually, diana ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC Equipment
The Bond does do ISH. In fact it utilizes the same detection kit so you have only to buy the probes, not the additional detection kit that could (depending on your volume of ISH requests) expire. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "Sheila Fonner" 2/25/2011 10:20 AM >>> Cindy, We used to use the Dako stainer, and we still have it as a back-up if necessary, but we have recently (in the last year) bought a Ventana Ultra. You can put 30 slides at a time on it, but you do not have to batch the slides. It is a continuous feed machine, which means that as soon as a slide is done, you can take it off and run something else. You do not have to have all of the slides from a case together side by side. It is bar-code driven and will find the slides no matter where you put them. Each slide drawer runs independently of the others. Ours has been wonderful. The technical assistance is fantastic also. They will help you to initially work up all of your antibodies. You can use theirs, or you can use third party antibodies and place them in a prep-kit. You can also use RTU or concentrates. It's really up to you. I would highly recommend it if you have a large volume. We demo'd the Biocare Intellipath also. I liked the machine, but it was really just a step up from the Dako. Leica has the Bond instruments, which a lot of people like, but for us, it didn't work because we wanted to be able to do ISH. Also, the Leica limits you to drawers of ten slides. So when you load a drawer, with 1 slide or 10, you can't use it again until that run is finished. Hope this helps. If you have any questions, you can contact me. Have a great day! Sheila -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cynthia Pyse Sent: Wednesday, February 23, 2011 2:37 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC Equipment Hi Histonetters I currently use a Dako stainer for my IHC staining. It is a work horse with very little problems. It is a older model that we may need to replace in the near future. What is everyone using out in histoland. I would be perfectly willing to purchase another Dako but I want to explore all avenues before making a decision. What are the pros and cons of the instruments any of you are using. How often is the machine down? What is the capacity? We run the Dako twice daily usually to the capacity of 48 slides. I would like to hear only from actual user of the instrumentation, no vendors please. This is only a fact finding e-mail. Thanks in advance for all your input. Cindy Cindy Pyse, CLT, HT (ASCP) Laboratory/Histology Supervisor X-Cell Laboratories 716-250-9235 e-mail cp...@x-celllab.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Any problems with Bond Max Heater Assembly (SSA)
Hi Folks, Someone from Leica should answer this question for you (Sara, are you listening in?), but I have a Bondmax and from time-to-time one of the heating pads will burnout. The other 9 positions continue to work. It used to be that the whole assembly was automatically replaced but now they manufacture them in such a way that an individual heater pad can be replaced. If you have a service contract this work will be covered. I think one or two heating pad errors per year could be expected - at least that has been my experience, but as I said, it doesn't slow us down much because the other 9 positions remain functional. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "Rathborne, Toni" 1/28/2011 11:17 AM >>> >>> We have a Bond III that's about a year old. I'm also interested in the answer. What does Leica say is the normal replacement period for this item? Is it covered under your service contract? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu]On Behalf Of Settembre, Dana Sent: Friday, January 28, 2011 10:13 AM To: 'Feher, Stephen'; Nancy Schmitt; histonet@lists.utsouthwestern.edu Cc: Delia, Catherine Subject: [Histonet] Any problems with Bond Max Heater Assembly (SSA) Importance: High Has anyone had any problems with the Leica Bond Max's Heater assembly. We have 2 Bonds that are less than 2 years old and one of the heater assembles went. That is 10 less slides that we have to work with. Thank you, Dana Settembre University Hospital - UMDNJ Newark, NJ 973-972-5232 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Feher, Stephen Sent: Friday, January 28, 2011 10:06 AM To: Nancy Schmitt; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] LEAN Processes/ new lab Hi Nancy, We built a brand new pathology lab from the bottom up last year. Drop me an email off line or give me a call and we can discuss our lessons learned etc. I worked with a LEAN workflow specialist on the design and equipment placement prior to the plans being approved. We included the architect in all of out discussions about this. You can always come to snowy New Hampshire and check it out yourself? Steve Stephen A. Feher, MS, SCT (ASCP) Pathology Supervisor Catholic Medical Center 100 McGregor Street Manchester, NH 03102 603-663-6707 sfe...@cmc-nh.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Nancy Schmitt Sent: Friday, January 28, 2011 9:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] LEAN Processes/ new lab Hi Histonetters- I am looking for input, pros and cons or anything else you can offer. We are designing a new histology lab to move to and I am interested in how others have handled this along with using LEAN processes. What would you do differently? What would you do the same? Are you in the tri-state (IA, WI, MN) area and open to visitors? Thank you in advance for your thoughts- Nancy Schmitt MLT, HT (ASCP) Histology Coordinator United Clinical Laboratories Dubuque, IA NOTICE: This email may contain legally privileged information. The information is for the use of only the intended recipient(s) even if addressed incorrectly. If you are not the intended recipient, please notify the sender that you have received it in error and then delete it along with any attachments. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE This message and any included attachments are from Somerset Medical Center and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or trade secret information entitled to protection and/or exemption from disclosure under applicable law. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call Somerse
Re: [Histonet] ihc slides
I should add that the DAB step may also have "quenched" all of the linked peroxidases even before the hematoxilin was added (which would finish off any that remained). Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "Cynthia Pyse" 1/24/2011 8:57 AM >>> Happy Monday Histonetters I need to pick the brains of the IHC experts out there in histoland. I run IHC on the Dako autostainer. On one of our runs there was not enough DAB to dispense on all the slides. My tech dehydrated the slides and cover slipped them. We printed the stain log on the run and confirmed that the DAB was not dispensed on three slides. The cover slips were removed and the slide were soaked in xylene to remove the cover slipping media. The slides were rehydrated and DAB was applied. There was still no reaction after 10 minutes, The DAB was reapplied for up to 30 minutes with no reaction. Can anyone out there give me a reason that the slides would not react after the reapplication of the DAB. I have done this before when the DAB solution was short on the run and it worked. I can't figure out why this didn't react. I called tech service at Dako and talked to the supervisor, she agrees with me that the reapplication should have worked. I checked the run log and the only missing step is the DAB. The antibodies with the missing DAB have their own controls on the slides. Any thought out there as to what the problem is. I have had the PM done on the Dako which should resolve the problem according to Dako. There has not been a problem since. This mystery will drive me crazy until I can figure out why. I could use help. Thanks in advance, I appreciate any input. Cindy Cindy Pyse, CLT, HT (ASCP) Laboratory/Histology Supervisor X-Cell Laboratories 716-250-9235 e-mail cp...@x-celllab.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] ihc slides
Cynthia, Hematoxylin will inactivate the peroxidase (link) in the tissues. You must go back and re-do the link step and hopefully the antigenic sites will still accept more streptavidin (or similar) conjugated antibody. Cheer. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "Cynthia Pyse" 1/24/2011 8:57 AM >>> Happy Monday Histonetters I need to pick the brains of the IHC experts out there in histoland. I run IHC on the Dako autostainer. On one of our runs there was not enough DAB to dispense on all the slides. My tech dehydrated the slides and cover slipped them. We printed the stain log on the run and confirmed that the DAB was not dispensed on three slides. The cover slips were removed and the slide were soaked in xylene to remove the cover slipping media. The slides were rehydrated and DAB was applied. There was still no reaction after 10 minutes, The DAB was reapplied for up to 30 minutes with no reaction. Can anyone out there give me a reason that the slides would not react after the reapplication of the DAB. I have done this before when the DAB solution was short on the run and it worked. I can't figure out why this didn't react. I called tech service at Dako and talked to the supervisor, she agrees with me that the reapplication should have worked. I checked the run log and the only missing step is the DAB. The antibodies with the missing DAB have their own controls on the slides. Any thought out there as to what the problem is. I have had the PM done on the Dako which should resolve the problem according to Dako. There has not been a problem since. This mystery will drive me crazy until I can figure out why. I could use help. Thanks in advance, I appreciate any input. Cindy Cindy Pyse, CLT, HT (ASCP) Laboratory/Histology Supervisor X-Cell Laboratories 716-250-9235 e-mail cp...@x-celllab.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] glycogen degeneration???
Hi folks, Has anyone experienced glycogen disappearing from previously "excellent" control blocks of liver. A glycogen-laden liver should be consistent throughout should it not? (ie we can't cut through it can we??). Or is oxidation a possible culprit here (not heard of it)? Really interested to get some feedback on this one. Thanks so much, Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Klotz fixative
The benefit of Klotz is it preserves tissues/organs while not altering the color and texture "much". Therefore good for demonstration labs. We used to store teaching specimens in Klotz in the walk-in cooler (+ 4C) to aid in the preservation even further. It is not adequate for long-term storage as far as I am aware. I have no experience with the affects on histological preparations. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> John Kiernan 2/1/2010 2:08 AM >>> A Google search for KLOTZ FIXATIVE brings up many hits, with indications that "Klotz solution" is an embalming fluid for gross anatomy. That's not the same as as a fixative. Follow up the links and see if they match what you need. Embalming fluids do not necessarily provide good histological fixation. John Kiernan Anatomy, UWO London, Canada = = = - Original Message - From: "Perry, Margaret" Date: Friday, January 29, 2010 16:00 Subject: [Histonet] Klotz fixative To: "histonet@lists.utsouthwestern.edu" > > Do you know of anyone that sells Klotz fixative? We use it > in a pre vet class but would like to eliminate the > choloralhydrate since it is such a hassel due to it's being a > controlled substance. Is there a substitute fixative that would > retain the properties of Klotz? > > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Advice from Canadian Labs
Hey Pam, No problem, I will. Nice to hear someone south of our border has heard of Canada's smallest province!! :-) Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> 1/18/2010 4:00 PM >>> It is s good question and since we are all moving that way I hope any advanced answers will be posted to all of us. PEI is a beautiful place!! Pam Marcum UAMS Sent from my Verizon Wireless BlackBerry -Original Message- From: "Greg Dobbin" Date: Mon, 18 Jan 2010 15:29:06 To: Subject: [Histonet] Advice from Canadian Labs Hello Canadian Colleagues, I am wondering what other clinical institutions in Canada are doing with regard to retention of hardcopy consult reports. Here in prince Edward Island we now have an electronic patient health record province-wide so we no longer have hardcopy surgical reports filed in the lab. Consult reports received from reference laboratories are being scanned into the patient's report and checked and verified for ledgibility. Any section that does not scan clear enough to read easily is edited to match the hardcopy. So in our lab, the patient report and any associated consult reports will be stored indefinately electronically. The CAP guidelines (which we refer to but are not held to) suggest "Surgical Consultation" reports should be kept indefinitely. I think therefore, we are meeting the expectation here, but how long should I retain the hardcopy of these consultation reports, 2 years? 20 years?? What are others doing in this regard? Thanks. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Advice from Canadian Labs
Hello Canadian Colleagues, I am wondering what other clinical institutions in Canada are doing with regard to retention of hardcopy consult reports. Here in prince Edward Island we now have an electronic patient health record province-wide so we no longer have hardcopy surgical reports filed in the lab. Consult reports received from reference laboratories are being scanned into the patient's report and checked and verified for ledgibility. Any section that does not scan clear enough to read easily is edited to match the hardcopy. So in our lab, the patient report and any associated consult reports will be stored indefinately electronically. The CAP guidelines (which we refer to but are not held to) suggest "Surgical Consultation" reports should be kept indefinitely. I think therefore, we are meeting the expectation here, but how long should I retain the hardcopy of these consultation reports, 2 years? 20 years?? What are others doing in this regard? Thanks. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] pH of 10% NBF
Hi Folks, If the pH of our 10% Neutral Buffered Formalin is reading 7.01 (recycled) and 7.12 (made from new formaldehyde), should I try to get to 7.2? If yes-how- add sodium bicarbonate? I look forward to your responses. Warm regards, Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Cutting Frozen Lung
Hi Meghan, Healthy lung is terrible to cut frozen. All those air spaces collapse on sectioning. Unhealthy lung on the other hand is a dream to cut! I have heard it suggested to perfuse the lungs with dilute frozen cutting compound (eg OCT or similar) prior to selecting the piece(s) for frozen sectioning. In theory it should work well but I have not tried it myself. Otherwise, a quick swipe of the cutting surface with your thumb just before sectioning might help a little (maximizing the amount of viewable tissue in the section) but your sections (of healthy lung)will never be "beautiful" in my experience. good luck. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> Meghan Tucker 1/4/2010 11:23:01 AM >>> Hello! I am looking for some helpful hints for cutting frozen lung tissue. I have tried just about everything I can think of! Thanks! ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Blade Dispenser Thingy
Hi Sally, Fisher is aware of the problem (or should be). I contacted Fisher Canada about it probably a year ago or so and had them replace 4 boxes for me. They made it seem as though this was a brand new phenomenon that they had not heard of before but I've known it to be a sporadic problem for years and I figured I couldn't have been the only one to have experienced this problem! They logged my complaint and forwarded the information on to the manufacturer (a Japonese company I believe). But as yet, I have't seen any design improvements to the dispensers. Maybe a Fisher person on this list would like to comment on whether the manufacturer has made any progress toward improving the dispensers. Cheers. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "Breeden, Sara" 12/15/2009 10:52 AM >>> I have Feather dispensers with blades that I have to pry out, which is probably not a good thing. I've tried freezing the dispenser, putting it in an oven for a while and whacking the darned thing on the floor (not necessarily all at the same time..). Have we had this discussion before and have we solved this problem? Help? Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] opinion please
I agree with Renee. It will proabaly be at least as costly to have to re-validate everything! Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> Rene J Buesa 12/7/2009 11:21 AM >>> Perhaps yes or perhaps no, that is the problem. >From now on you cannot know if something that did not work was because of the >fridge malfunction. Advise? (Costly advise?): discard everything and get an alarm system for your fridge after you repair it (or get a new one).. René J. --- On Mon, 12/7/09, Kim Merriam wrote: From: Kim Merriam Subject: [Histonet] opinion please To: "Histonet" Date: Monday, December 7, 2009, 9:51 AM Help! My refrigerator died over the weekend. This fridge had most of my antibodies and IHC reagents. When I came in, the temperature inside the fridge was a balmy 37C! Do you think any of my reagents are still usable? Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Morgue topic: Chain of Custody for personal effects
Hi Folks, Can any of you share with me your procedure for ensuring chain of custody for personal effects of decedents. Along with documenting a clear procedure for all stakeholders to abide by, I also need to solve some the practical aspects like what to use for a tamper-resistant bag for the effects and should I have a 3-part form (1 copy for us, 1 for the Commissionaire and 1 the funeral home) that lists all the personal items, money etc. Jewellery is only removed if there is a risk of damage (eg wrist watches, necklaces). Piercings are generally not removed. Rings are usually taped. A little background info: Most of the decedents we receive in our morgue are "at the back door", usually via ambulance (our coroner system does not have their own vehicles) or from the ER. Occasionally we will get a hospital case from the nursing floors. The Commissionaire (security) receives the bodies and unlocks the morgue for delivery. The pathologist on call is notified and an autopsy is scheduled. After the autopsy we inform the Commissionaire that the remains have been released and he/she in turn notifies the funeral home and looks after signing the remains over to the funeral home staff. Thank you! Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Leica Bond/Novocastra Reagents
Almost always receive on time. I can think of only one instance in a year and a half where anything was on back order (it was an antibody). Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> Paula Lucas 10/5/2009 12:20 PM >>> Hello, We are considering the Leica Bond and their reagent rental or acquisition option, and I have a question regarding back orders. If anyone places an order through Leica for their Bond reagents (novocastra), are there any problems with back orders on antibodies, detection system and other reagents needed to run the Bond? Or, do you usually get the items right away? Thank you, Paula Lucas Lab Manager Bio-Path Medical Group Fountain Valley, CA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] paraffin sectioning static
Heavy breathing- the pathologist walking by might do a double-take, but the humidy from your breath should get rid of immediate static. Alternatively use a humidifier in the lab, but of course that comes with its own set of problems (ie aerosolizing Pseudomonas and other environmental bacteria and/or fungii). Just a thought (and a little humour for good measure!). Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> Emily Sours 10/1/2009 4:05 PM >>> Any tips on getting rid of static when paraffin sectioning on a microtome? It's so insanely bad, the block is picking up the ribbon as it goes back up every third section. Emily ...the thrill of being close to that hidden knowledge. That's the way I feel when I read Nabokov. Encrypted within his words, encoded indecipherably, ambiguously, is the equivalent of the secret of lightning. Something akin to the secret code of higher human consciousness, the DNA, the genome of genius. -Ron Rosenbaum ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] mib-1
"Alcohol based" suggests to me an absence of formalin. In which case, epitope retreival is not indicated. Also, you didn't say if it had been working and stopped or if this was a new marker for your lab. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> 9/16/2009 3:44 PM >>> I have to turn to the experts here.? I am having problems with the MIB-1.? I am having an issue with the patient samples working.? We use an alcohol based fixativeit has always worked until recently. I have tried everything I can think of to no avail.? I have decreased the titer, increased the incubation, tried a different retrieval solution, tried a different detection...nothing is working. Anyone else having any issues? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Best IHC stainer
I echo Joyce's comments! Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "Weems, Joyce" 8/26/2009 10:57 PM >>> They are both good instruments. From my experience pricing is high for the Ventana reagents and there seems to be an increase in DAKO pricing from what I see on the Net. Have you looked at the Leica Bond? We found that to be a good fit for us. Started out to add it to our DAKO line, but ended up changing completely. It also does ISH, and is a continuous feed - up to 10 slides at a time. Good luck, Joyce -Original Message- From: histonet-boun...@lists.utsouthwestern.edu on behalf of Gareth Blaeuer Davis Sent: Wed 8/26/2009 8:04 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Best IHC stainer The lab I work in has been doing demos on IHC stainers Ventana Benchmark XT and Dako's Autostainer Plus. We could really use feed back from current users of both. We are having a hard time deciding between the two, so any input would be great. What are the Pros and Cons with both. Thanks, Gareth Blaeuer Davis, B.S., HT _ Hotmail® is up to 70% faster. Now good news travels really fast. http://windowslive.com/online/hotmail?ocid=PID23391::T:WLMTAGL:ON:WL:en-US:WM_HYGN_faster:082009___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. Déclaration de confidentialité Le présent message (y compris les annexes) peut contenir des renseignements confidentiels à l'intention d'une personne ou d'un organisme particulier. Si vous avez reçu la présente communication par erreur, veuillez en informer l'expéditeur immédiatement. Si vous n'êtes pas le destinataire prévu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer immédiatement de votre système informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] STD testing in Lab
Oh my gosh! You're right. My day is suddenly not so drab!! LOL Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "Blazek, Linda" 6/25/2009 3:48 PM >>> NO!! It's THURSDAY afternoon isn't it? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Greg Dobbin Sent: Thursday, June 25, 2009 2:20 PM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] STD testing in Lab Well I would check the local human rights laws. I'm not so sure your staff are going to be too thrilled with being tested for all these diseases! .I'M KIDDING of course. I'm sorry I can't contribute anything worthwhile (other than some comic relief) on what is otherwise a rather drab Wednesday afternoon. Cheers! Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> Jill Cox 6/25/2009 2:52 PM >>> Hi Histonetters!!We are trying to bring in house STD testing in our lab and would like to know what type of equipment everyone uses and feedback. We would like to take on HPV, GC Chlamydia and gonorrhea. Any information would be greatly appreciated!! Thank you.. Jill Cox HT (ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] STD testing in Lab
Well I would check the local human rights laws. I'm not so sure your staff are going to be too thrilled with being tested for all these diseases! .I'M KIDDING of course. I'm sorry I can't contribute anything worthwhile (other than some comic relief) on what is otherwise a rather drab Wednesday afternoon. Cheers! Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> Jill Cox 6/25/2009 2:52 PM >>> Hi Histonetters!!We are trying to bring in house STD testing in our lab and would like to know what type of equipment everyone uses and feedback. We would like to take on HPV, GC Chlamydia and gonorrhea. Any information would be greatly appreciated!! Thank you.. Jill Cox HT (ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Leica CV5030
Hi Patsy,
I will concur with Hazel but would add that the coverslipper has only
been hassle free for us as long we are using the better quality (ie
"Premium" grade) coverslips. Cheaper coverslips are not economical when
one considers the time lost to fix and reset the instrument and in some
cases re-coverslip slides. But since I switched to using the premium
grade year round (as opposed to only in the more humid months, which is
what I was doing to try andsave money!) I have had almost no glitches at
all!
Cheers!
Greg
Greg Dobbin, R.T.
Chief Technologist, Anatomic Pathology
Dept. of Laboratory Medicine,
Queen Elizabeth Hospital,
P.O. Box 6600
Charlottetown, PEC1A 8T5
Phone: (902) 894-2337
Fax: (902) 894-2385
"I find that the harder I work, the
more luck I seem to have."
- Thomas Jefferson
>>> "Horn, Hazel V" 6/23/2009 11:16 AM >>>
We absolutely love our CV5030. I really can't think of anything
negative to say about it.
Hazel Horn
Hazel Horn, HT/HTL (ASCP)
Supervisor of Histology
Arkansas Children's Hospital
1 Children's WaySlot 820
Little Rock, AR 72202
phone 501.364.4240
fax501.364.3155
visit us on the web at:www.archildrens.org
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Patsy
Ruegg
Sent: Monday, June 22, 2009 2:53 PM
To: 'Histonet'
Subject: [Histonet] Leica CV5030
I am looking for experiences with this coverslipper from Leica CV5030
model, good or bad.
Thank you,
Patsy
Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pru...@ihctech.net
web site www.ihctech.net
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RE: [Histonet] Cracked or shattered LN sections
I am positive no freezing spray being used. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "Sebree Linda A" 04/27/09 7:36 PM >>> Greg, Are you sure no one is using freezing spray when sectioning? The effect of freezing spray on the block would yield the appearance you describe. From: histonet-boun...@lists.utsouthwestern.edu on behalf of Greg Dobbin Sent: Mon 4/27/2009 2:04 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Cracked or shattered LN sections Hi Folks, Some lymph node sections (usually bigger ones-maybe almost as big as a dime) have a shattered appearance on the block surface. Subsequently the sections look like shattered glass or a mosaic of tissue pieces making up the whole. I think this is happening at the embedding station from trying to make sure that the tissue is flat in the mold (ie pressing the tissue down). Has anyone else out there have any similar experience with this problem? Is the solution as simple as cutting smaller pieces into the cassette at the time of trimming? Thanks. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Cracked or shattered LN sections
Hi Folks, Some lymph node sections (usually bigger ones-maybe almost as big as a dime) have a shattered appearance on the block surface. Subsequently the sections look like shattered glass or a mosaic of tissue pieces making up the whole. I think this is happening at the embedding station from trying to make sure that the tissue is flat in the mold (ie pressing the tissue down). Has anyone else out there have any similar experience with this problem? Is the solution as simple as cutting smaller pieces into the cassette at the time of trimming? Thanks. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Recycled formalin
Yes. ?? Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "Richard Cartun" 04/07/09 7:30 PM >>> Is anyone using recycled formalin for primary fixation of either surgical or autopsy tissue? Thanks. Richard Richard W. Cartun, Ph.D. Director, Histology & Immunopathology Director, Biospecimens Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Strange circles in IHC slides
Hi V. That my friend is the result of a air bubble on the section. If it had been unremoved xylene the hematoxylin would have been absent as well. Cheers. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "V. Neubert" 4/2/2009 1:12:18 PM >>> Hello, I really have some real links to real pictures of real relevance of the real histonet list. Really promised. http://img12.imageshack.us/img12/8513/ts0402162049.jpg http://img13.imageshack.us/img13/6514/ts0402162104.jpg So, has ever anyone experienced sth. like this? My conjugate control (every step except the antibody) was fine, nothing to be seen about DAB and no circles at all. I used Shandon single-use coverplates, sterile buffer, fresh antibody aliquots. Any idea? Thanks, V. Neubert ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Commercial Eosin
Hi Vanessa, Yes! I have our eosin staining down to just 10 secs! I am considering just letting the slides hover over top of the eosin for a minute instead of actually dipping! LOL Very strong indeed. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "Vanessa J. Phelan" 3/26/2009 3:23:49 PM >>> Hi everyone, Has anyone had the experience of massive overstaining with commercial Surgipath Alcohol-based Eosin? I manually stained with Surgipath Hx for 2mins, then bluer the eosin for 30secs and the whole tissue was completely pink! Thanks V ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC stainer
I guess I would have to echo that as well! Cheers! Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "Houston, Ronald" 3/13/2009 4:45:14 PM >>> Can't speak highly enough of the BondMax form Leica Microsystems. If cost is an issue and you're looking at what to avoid, IMHO avoid anything Ventana Ronnie Houston Anatomic Pathology Manager Nationwide Children's Hospital Columbus OH 43205 (614) 722 5450 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Stephanie Weaver Sent: Friday, March 13, 2009 3:17 PM To: histonet post Subject: [Histonet] IHC stainer I am in a veterinary diagnostic lab. In the past we have had very few requests for IHC and have always sent slides out to another lab to perform IHC as needed. It is time for us to start doing our own and join the modern age. We have several certified technicians, but none have experience with IHC and we typically have a relatively high turnover rate. Therefore, I am hoping to be able to buy an automated stainer. In the past most people on the list seemed to be very happy with the Dako autostainer, but this past week has brought so many bad remarks about Dako's service that I am reconsidering. We probably will not need a high capacity autostainer, but I would like walk-away capability with an easy to use system. It will need to accept other companies reagents, since veterinary infectious disease antibodies aren't often sold by the major companies. Also, cost is an issue and I would like to be able to bargain shop for reagents through other companies. Does anyone have any recommendations, or warnings as to what to avoid? In a related issue, where do other animal tissue people get their antibodies for infectious diseases, e.g. Parvovirus, canine distemper, or FIP? Thanks for the advice! Stephanie Weaver Texas Veterinary Medical Diagnostic Lab ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Confidentiality Notice: The following mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. The recipient is responsible to maintain the confidentiality of this information and to use the information only for authorized purposes. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that any review, use, disclosure, distribution, copying, printing, or action taken in reliance on the contents of this e-mail is strictly prohibited. If you have received this communication in error, please notify us immediately by reply e-mail and destroy all copies of the original message. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC stainer
I echo everything Julie said! Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> Julie Trejo 3/13/2009 4:41:06 PM >>> I suggest using the Leica Bond-Max. I, an HT(ASCP)cm, did not have any IHC experiance, but the Bond-Max makes it happen. Just putting dried slides on, patient info in the computer, slides labeled and then in 3-4 hours, it's all done. Just dehydrate and clear. Very simple and now giving me time to learn IHC and troubleshooting. Some of the antibodies come "Ready to use" from Leica, and we titer others that aren't "Ready to use" (Biocare, Dako etc). I've seen co-workers in the past struggling with immuno's using the Dako, and taking alot of time and still counter-staining by hand. The Bond-Max allows us (just three of us) to do all routine H&E's, special stains, recuts, deepers and Immuno's in one day. Not bad. Julie On Fri, Mar 13, 2009 at 2:16 PM, Stephanie Weaver wrote: > I am in a veterinary diagnostic lab. In the past we have had very few > requests for IHC and have always sent slides out to another lab to perform > IHC as needed. It is time for us to start doing our own and join the modern > age. We have several certified technicians, but none have experience with > IHC and we typically have a relatively high turnover rate. Therefore, I am > hoping to be able to buy an automated stainer. In the past most people on > the list seemed to be very happy with the Dako autostainer, but this past > week has brought so many bad remarks about Dako's service that I am > reconsidering. We probably will not need a high capacity autostainer, but I > would like walk-away capability with an easy to use system. It will need to > accept other companies reagents, since veterinary infectious disease > antibodies aren't often sold by the major companies. Also, cost is an issue > and I would like to be able to bargain shop for reagents through other > companies. Does anyone have any recommendations, or warnings as to what to > avoid? > > In a related issue, where do other animal tissue people get their > antibodies for infectious diseases, e.g. Parvovirus, canine distemper, or > FIP? > > Thanks for the advice! > > > > > Stephanie Weaver > Texas Veterinary Medical Diagnostic Lab > > > > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Julie Trejo, HT(ASCP)cm Saint Louis University Department of Dermatology 314-256-3413 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] silly question about goat serum
Hi Emily, Normal as opposed to immune serum. If a goat outside of a "Specific Pathogen Free (SPF)" facility were to actually have no Ab's it would be far from normal! I'm picturing the Boy in a Bubble episode on Seinfeld- only substitute one kid for another!! LOL (or groan if you prefer). Enjoy the day folks! Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> Emily Sours 3/12/2009 11:41:30 AM >>> What exactly does non-immune goat serum mean? The goat hasn't been exposed to any specific antigen, therefore it's normal? Emily -- prometheus, thief of light, giver of light, bound by the gods, must have been a book. -mark danielewski, house of leaves ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Embedding bone marrow aspirates
Hello Colleagues, I'm looking for opinions/experience regarding the following idea: I am thinking about using Histogel (not pushing this product over another, just don't know what else to call it!!) to localize the aspirate to a more confined area, hopefully more consistently on a single plane, and perhaps improve the "cutability" of the more brittle, crumbly specimens. Currently we filter BM aspirates with a piece of lenspaper, scrape the tissue from the "filter" onto a smaller piece of lenspaper that is then folded and placed between 2 biopsy pads in a cassette. Then the embedder has to scrape the processed material into the embedding mold and hopefully get most of it in the same plane (which he does reasonably well). What I would like to do is scrape most of the tissue to the bottom of the lenspaper filter cone, add histogel and then place the resulting pellet in a cassette for subsequent processing. I look forward to reading your knowledgeable replies. Have a nice weekend everyone. Sincerely, Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] BVD Antibody
Did you check VMRD? http://www.vmrd.com/products/antibodies/detailInfIG.aspx?CATNO=D89 Bovine Viral Diarrhea Virus MAb (gp55) Catalog No.: D89 Description: Binds to 55 kDa of BVDV. Made using NADL strain. Binds to most BVDV strains. Does not bind Oregon C24V strain. IgG2a isotype. Suitable for immunofluorescence, immunohistochemistry and virus neutralization. This monoclonal antibody is produced as mouse ascites fluid, clarified by centrifugation, and filtered through a 0.2 micrometer filter. The concentration is 1.0 mg/ml in phosphate-buffered saline, preserved with 10 ppm ProClin 300. Format: Mouse ascites Size: 0.1, 0.5 and 1 mg Expiration: One year from date of QC release. Storage: Store at 2-7°C. Do not freeze! Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. Déclaration de confidentialité Le présent message (y compris les annexes) peut contenir des renseignements confidentiels à l'intention d'une personne ou d'un organisme particulier. Si vous avez reçu la présente communication par erreur, veuillez en informer l'expéditeur immédiatement. Si vous n'êtes pas le destinataire prévu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer immédiatement de votre système informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Glass coverslippers
The CV5030 works very well for us-as long as we spend a little more buying the higher grade coverslips that don't tend to be affected by humidity as much. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IF on cell lines
Hi April, This may not be useful to your colleague at all, but here goes: I used to place a round glass sterile coverslip in the bottom of each well of a 24-well culture plate before seeding and then grew the cells on the coverslip. When ready the coverslip was removed (using a hyperdermic needle with the bevelled tip bent backwards to catch the edge of the cs). The coverslips were then picked up with "eyelash" forceps and placed in a little porcelain coverslip rack and placed in cold acetone (-20 C) to fix for 10 mins. Following IF staining the round coverslip was mounted on a regular coverslip which was in turn mounted on a glass slide for viewing. In case you haven't already figured this out, acetone melts the plastic chamber slides. My coverslip method, while effective, is obviously much more laborious (which I suppose is why the chamber slide was developed in the first place! LOL). Hopefully the 4% paraform. works. Is methanol fixation an option?? Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> Aprill Watanabe 2/16/2009 9:13 AM >>> I have a colleague who is trying to do IF on cell culture cell lines. She is growing them on cell culture treated chamber slides and fixing with 4% formaldehyde. Her problem is that after fixation and a few washes all the cells are detaching. Any suggestions? I think she is going to try 4% paraformaldehyde first as someone has already switching to that. Aprill Watanabe, B.S. Research Associate Integrated Cancer Genomics Division Tissue Microarray Center (TMA) Translational Genomics Research Institute (TGen) main: 602-343-8822 Fax: 602-343-8840 awatan...@tgen.org www.tgen.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC and ISH
Jean, The Bond Max is different than its main competitor in that the Bond uses the same detection kit for both IHC and ISH. One just has to buy the fluorocein reagent for the ISH method. So all you have is one reagent to worry about expiring rather than an entire detection system. Other instruments (they know who they are) require you to buy an entire separate detection system to do ISH and so if your volumes for ISH are low, you will waste a lot of money in expired kits. Cheers. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> "Taylor, Jean" 2/2/2009 1:50:41 PM >>> Hi everyone, I am currently in the process of evaluating IHC equipment to replace my old Dako Autostainers. The pathologists I work for are interested in bringing ISH into our lab. My questions are, what IHC instrument have you found works the best for IHC and ISH, and what kind of training did your personnel go through to become qualified to perform ISH? Thank you! Jean Taylor, HT(ASCP)QIHC ICH Tech Meriter Labs Madison, WI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Cytology Autostainers
Hi Folks, What's out there? What's hot and what's not? Anyone using their cytology autostainer for histology special stains? Welcome all input (including vendors). Cheers! Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] double staining on the ventana XT
Hi Jennifer, Your Ventana Application Specialist should be more than willing to help you thru this process. I demo'd an XT last year and I recall double-staining was very straight forward. Unfortunately the details of which were not retained by me! Good luck. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> 01/21/09 8:41 PM >>> Greetings, We are looking into doing some immuno double staining. I searched the archives and found a few similar inquiries but no responses. Anyone with experiences to share I would love to hear what you have to say. Everything from what not to do to what works well for you. Many Thanks, Jennifer - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Auto-IHC Staining Issues
Hi Paula, You really have to tell us which instrument you are using to get a concise answer. Other details like how the slides are dewaxed for instance would also definately help. Regards, Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson >>> Paula Lucas 01/21/09 8:04 PM >>> Hello, I've been having a problem with the staining quality from an auto-IHC stainer and I was wondering if any of you experience the same thing. I notice sometimes that the staining in the tissue isn't complete, like there are areas where the fluid (one or more of the steps involved) didn't get in contact with the tissue, thus resulting in patchy, skipped staining. I'm thinking that this must be a dispensing problem. Is this a general thing with all auto stainers - ones like DAKO or Lab Vision's stainer or Biocare Medical's stainer? These stainers dispense fluid onto the slide. I hope this explains things and I really hope that I can get some feedback. It's getting very frustrating. Thanks in advance, Paula Lucas Lab Manager Bio-Path Medical Group Fountain Valley, CA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Dako and Leica immunostainers
Sally, I think you answered your own question! You wrote: "If so, why would someone want one of these stainers?" And yet, they sell so many! I have found the Bond to be an excellent instrument and in my opinion it is unrivalled in the market place. I have just recently spent 10 months doing extensive evaluations of Bond, Dako and Ventana. I brought each instrument in-house to try. When all was said and done the Bond was the best fit for my lab. Each lab needs to evaluate what is the best fit for their particular situation (ie number of staff, experience level of staff, volume of tests, workflow, etc.). But with regard to the cost for online dewax and retreival: what is your time worth? Doing these steps off-line means more tech time, more opportunity for mistakes and opens the possibility of peripheral instrumentation failing and holding everything up down the line. As for the cost per slide: I spent many agonizing hours going over pricing info for all 3 instruments (trying to "uncode" the sales speak) and in the end (by my calculations) the difference between 2 of the 3 was less than a dollar per test and the 3rd was considerably higher. I won't say which one is which, but I will say the Bond was not the higher of the 3. Hope this has helped. Cheers! Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "And in the end it's not the years in your life that count. It's the life in your years." - Abraham Lincoln >>> "Sally Price" <[EMAIL PROTECTED]> 11/20/2008 7:35 PM >>> All, After reading this thread I just had offer my comments. I'm not a big fan of systems that do the dewax and AR, primarily because it costs way too much to automate these steps. I've never used the Bond, but I hear that you've gotta put some plastic thingy - that probably costs too much - on top of each slide, you gotta use their detection reagents - which probably cost more than other companies, they charge you for empty barcoded reagent containers, all the slides in the same tray have to use the same detection reagents - which means that the continuous-feed feature has some serious limits, it can't do double-stains, and they have less than 50 IVD-approved antbodies. Can someone verify for me if all this issues are true? If so, why would someone want one of these stainers? The Dako stainer is a dinosaur and with all the newre/better ones available, they should probably take it off the market. Cheers, Sally -Original Message- From: Josie Britton <[EMAIL PROTECTED]> To: Michael Bradley <[EMAIL PROTECTED]>; Histonet@lists.utsouthwestern.edu Sent: Tue, 18 Nov 2008 11:31 am Subject: RE: [Histonet] Dako Hi, We just got the new Bond Max IHC stainer and we love it. You just cut the slides dry them and place them on the Bond. It does retrieval, antibody staining, and counter stain. You just dehydrate , clear, and mount your coverslip. It is easy to use. It has 3 individual slide tray's of 10. You can load more slides on the empty tray's and start a new batch while the others are running. We run into the pathologist's adding more antibodies to the list an hour after we have run the first batch frequently, so this feature is great. When you add more IHC's the run time on all the slide tray's run times do increase, but it's better than having to wait another 2-3 hours to put your next set of immuno's on. Hope this helps! Josie Britton HT Cheshire Medical Center 580 Court Street Keene, NH 03431 -Original Message- Message: 9 Date: Wed, 19 Nov 2008 17:17:52 -0500 From: [EMAIL PROTECTED] Subject: Re: [Histonet] Dako To: [EMAIL PROTECTED], [EMAIL PROTECTED], Histonet@lists.utsouthwestern.edu Just another FYI... The Biocare IntelliPath does the same thing, but has 4 trays of 10 and it allows you to make one of the trays?RUSH.? SO depending, on the volume?of IHC you do either of these machines would work out great. Roxanne ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d&#x
RE: [Histonet] Presidential Voting Infomation
I don't remember seeing this persons name on the Histonet before. Could he be a Republican Party volunteer who joins various listservs for the sole purpose of garnering support for Mr. McCain?? It certainly would not surprise me. The fact that so many of us have read it already tells me he's accomplished his mission! Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "And in the end it's not the years in your life that count. It's the life in your years." - Abraham Lincoln - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] C-erbB-2 SP3 clone
Hello Folks, Is anyone out there using this clone from LabVision? I am trying to decide what volume to purchase in order to try the Ab out. There is a 0.1 mL size available and that would be my preference as long as the starting dilution is sufficiently high to allow several validation runs. So my question is what dilution range am I looking at for this clone? Thank you. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "And in the end it's not the years in your life that count. It's the life in your years." - Abraham Lincoln - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Paper (teabag) Bx bags-Source?
Hi Folks, The archives seem to be temporarily(?) unavailable. Can someone point me in the direction of a supplier of the paper teabag type of Bx bags? Thank you. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PEC1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "And in the end it's not the years in your life that count. It's the life in your years." - Abraham Lincoln - Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ? l'intention d'une personne ou d'un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer imm?diatement de votre syst?me informatique. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
