[Histonet] FISH signals
Hi, how the FISH signals in FFPE tissue can be strenghten? by standard procedure only weak signals for EGFR can be obtained, so far I tried different time in Vysis protease 1 (45, 60, 70 min), for other probes 70 min works quite well on our FFPE, but not for EGFR? what other modifications can I try? many thanks Irena ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] cryostat
Hi,I wonder what is the way of removing shavings/trimmings from the cryostat in other lab?, with the wet paper? gauze?, household vacuum cleaner - yes I saw this in one lab!?thanksIrena ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Urine cytologies
we prefer filtration of urines through 5 mikron policarbonate filters Irena Kirbis Ljubljana, Slovenia > To: bryan.wat...@neiurology.com > From: dkb...@chs.net > Date: Tue, 25 Oct 2011 09:46:06 -0400 > Subject: Re: [Histonet] Urine cytologies > CC: histonet@lists.utsouthwestern.edu; > histonet-boun...@lists.utsouthwestern.edu > > Saponin and Calcium Gluconate. Let me know if you would like the > procedure and I will send it to you under separate cover. > > Debbie M. Boyd, HT(ASCP) l Chief Histologist l Southside Regional Medical > Center I > 200 Medical Park Boulevard l Petersburg, Va. 23805 l T: 804-765-5050 l F: > 804-765-5582 l dkb...@chs.net > > > > > > > > Bryan Watson > Sent by: histonet-boun...@lists.utsouthwestern.edu > 10/25/2011 09:26 AM > > To > "histonet@lists.utsouthwestern.edu" > cc > > Subject > [Histonet] Urine cytologies > > > > > > > What is a good reagent for lysing red cells in extremely bloody urine > specimens for cytology? > thanks, > Bryan > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > -- > Disclaimer: This electronic message may contain information that is > Proprietary, Confidential, or legally privileged or protected. It > is intended only for the use of the individual(s) and entity named > in the message. If you are not an intended recipient of this > message, please notify the sender immediately and delete the > material from your computer. Do not deliver, distribute or copy > this message and do not disclose its contents or take any action in > reliance on the information it contains. > ___ > 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] Paraffin blocks from cell line
we made a cell block from cell line by following procedure: - centrifuge cell suspension - fix the sediment overnight in formalin (for tissue fixation) - optional, the button will be anyway fixed by subsequent procedure - centrifuge and decant the formalin - add few drops of liquid agar (not too hot) - as the button solidify you can put it in cassette and process as diagnostic tissue. We got good immunoreactions on sections prepared from melanoma cell line for example. hope this helps Irena Kirbis > From: kst...@mcw.edu > To: histonet@lists.utsouthwestern.edu > Date: Tue, 17 May 2011 13:45:41 -0500 > Subject: [Histonet] Paraffin blocks from cell line > > Hi histonetters, > > Has anyone made up paraffin blocks from cells from a cell line. This is new > to me. I assume they will be in some kind of a media. We will be getting the > cells and incubating them to grow. > Thanks in advance for the help > > Kathryn Stoll, HT(ASCP) > Depatment of Pathology > Medical College of Wisconsin > 9200 W Wisconsin Ave > Milwaukee WI 53226 > 414.805.1525 > kst...@mcw.edu > > ___ > 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] ArrayMold System
Hi, we have Arraymold more than a year now and we prepared around 10 TM's with it, it is really simple, easy and effective, however we found instructions to keep contructed TM overnight on 37 degrees uneffective since recipient and donor paraffin were not blend enough, we keep our TM's overnight on 40 degrees and next day half an hour on 60 (in moulds) if necessary we even prolonge this phase, this also help us to achieve that shorter tissue bars come down to the level. regards Irena Kirbis > Date: Tue, 8 Mar 2011 10:59:46 -0500 > From: alietee_langsd...@dfci.harvard.edu > To: histonet@lists.utsouthwestern.edu > Subject: re: [Histonet] ArrayMold System > > Hi, > I am also interested in hearing about the ArrayMold vs Beecher vs any other > do-it-yourself TMA systems. Any favorites systems/recommendations or > suggestions? > Thanks > ~Ally > Senior Research Technician > Medical Oncology > Dana-Farber Cancer Institute > re: > Wondering if anyone has used/purchased the ArrayMold "tissue micro array > system"?? Positive and negative feedback would be greatly appreciated :) > > Nancy Heath, HT(ASCP) > > March 10, 2011 is Histotechnology Professionals Day > > > Senior Research Technician > Medical Oncology > Dana-Farber Cancer Institute > > > > > > The information in this e-mail is intended only for the person to whom it is > addressed. If you believe this e-mail was sent to you in error and the e-mail > contains patient information, please contact the Partners Compliance HelpLine > at > http://www.partners.org/complianceline . If the e-mail was sent to you in > error > but does not contain patient information, please contact the sender and > properly > dispose of the e-mail. > ___ > 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] infectious fixed tissue and cells
Dear all, could anyone out there provide a scientific reason for wearing gloves handling slides with FFPE sections and FIXED cytology slides? I could understand that by wearing gloves we could prevent epithelial contamination of cytology slides but I would like to know which microorganism could survive FFPE procedure or cytology fixation and crowl around the slide after that? if cutting of FFPE tissue requires gloves what about archiving them, is it also necessary to wear the gloves? isn't the aerosol during the cutting more dangereous? are FFPE tissues and FIXED cytology slides considered infectious as the fresh tissue/cells? is there any study supporting practice of wearing the gloves or it is just superstition? or perhaps the gloves industry support? many thanks for your thoughts! Irena Kirbis ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] fixed cytology slides
Greetings to all, so far we handled wet-fixed cytology slides as health risk free without gloves, however there is no data regarding this on the net (or at least I wasn't able to find it). I would appreciate response on safety policy in other cytology labs regarding fixed cytology slides and/or any reference on the subject. many thanks Irena Kirbis Institute of Pathology Ljubljana, Slovenia ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Thyroid Smears
I guess that you're staining these fix slides by Papanicolaou?, perhaps the smears are too thick?, we process daily different kind of smears and always prepare one fix and one air dry smear for Pap and Giemsa staining and occasionaly we have an issue only very muccous samples which can coming off but never bloody smears except if they are very thick. one simples way to get rid of blood from cell suspension is a filtration through nylon mesh with 20 microns pores (published in Cytopathology - Heamorrhagic cytology samples- how to get best out of it) Irena > From: fawn.bo...@halifaxregional.com > To: histonet@lists.utsouthwestern.edu > Date: Wed, 12 Jan 2011 09:33:06 -0500 > Subject: [Histonet] Thyroid Smears > > Hello Everyone! > > Happy New Years to all! I have a question regarding the preparation of > thyroid smears. As of right now, we go up to the room and collect the thyroid > sample. The Pathologist makes the smears in the room and immediately puts > them into 95% Isopropanol to fix. We then complete the stain later on in the > day. The problem that we are encountering is that all of the blood and cells > are coming off of the slides before we make it through the entire stain. Does > anyone have any suggestions or are willing to share the procedure that they > use? We had a couple of suggestions that we recommended to the Doctor but > they were dismissed. I don't want to tell what are suggestions were so that > the doctor cannot accuse us of influencing every one else's opinions. > > Thank you in advance, > Fawn > - > This electronic message may contain information that is > confidential or legally privileged. It is intended only > for the use of the individual(s) and entity named as recipients > in the message. > > If you are not an intended recipient of this message, please > notify the sender immediately and delete the material from any > computer. Do not deliver, distribute, or copy this message, and > do not disclose its contents or take any action in reliance on > the information it contains. > > 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
RE: [Histonet] ArrayMold System
so far we have good experiencies with it, it's very easy to use eg. it's easy to prepare and construct TM, however we still struggle how to handle prepared TM in order to keep all punches on the cut sections, it seems that this part request some additional adjustment - at what temperature and how many cycles are needed to assure stabile TM, and of course it seems that cutting is crucial too. last but not least it's much much cheaper than Beecher, so for the labs just introducing and trying out TM in practice this way is better Irena > From: nhe...@lifespan.org > To: histonet@lists.utsouthwestern.edu > CC: histonet-boun...@lists.utsouthwestern.edu > Subject: [Histonet] ArrayMold System > > Wondering if anyone has used/purchased the ArrayMold "tissue micro array > system"?? Positive and negative feedback would be greatly appreciated :) > > Nancy Heath, HT(ASCP) > > March 10, 2011 is Histotechnology Professionals Day > > ___ > 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] RE: Edge affect on TMA?
how to prevent spots coming off during sectioning TM's? we do sometimes facing problems sectioning TM's, they're just falling apart? thanks for any hints Irena > From: hfe...@jhmi.edu > To: sette...@umdnj.edu; ahut...@dh.org; histonet@lists.utsouthwestern.edu > Date: Tue, 21 Dec 2010 16:13:31 -0500 > CC: > Subject: [Histonet] RE: Edge affect on TMA? > > Hello, We do not usually see that. If we do its very rare because I can't > think of any examples off hand. The usual problems are the spots came off or > shifted/smushed, not an edge effect. > > Helen > > > > > -Original Message- > From: histonet-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Settembre, > Dana > Sent: Tuesday, December 21, 2010 3:00 PM > To: 'Hutton, Allison'; histonet@lists.utsouthwestern.edu > Subject: [Histonet] Edge affect on TMA? > > Does anyone experience edge affect on TMA? > Dana Settembre > > ___ > 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
RE: [Histonet] carnoys fluid
I would recommend methanol or ethanol for fixation, smears itself can be culprit of inconsistent staining Irena > To: histonet@lists.utsouthwestern.edu > From: heather.mcl...@webmail.co.za > Date: Wed, 24 Nov 2010 19:42:51 +0200 > Subject: [Histonet] carnoys fluid > > Dear Histonetters, > > We do IHC on pap stained cyto smears. The results do not always "behave" > accordingly on the smears that have been exposed to Carnoys. Does anybody know > whether Carnoys Fluid affects IHC on cytology smears? I have seen papers that > suggest improved IHC after Carnoys but maybe Histonetters feel difefrently? > Thanks > Heather > > > South Africas premier free email service - www.webmail.co.za > > Free SMSs everyday! Gratis, Forniet! http://clients.wm.co.za/20030688/ > > > > ___ > 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] RE: Saponin Technique
for removing erythrocytes from hemorrhagic cytology samples we use filtration of bloody suspension through membrane filter with pore size 20 mikrons, you can find an article in Cytopathology. 2007 Jun;18(3):175-9. Epub 2007 Mar 27. it is worth trying! regards Irena > From: tmcne...@lmhealth.org > To: valerie.han...@parrishmed.com; histonet@lists.utsouthwestern.edu > Date: Mon, 25 Oct 2010 11:52:31 -0400 > CC: > Subject: [Histonet] RE: Saponin Technique > > I don't know about the procedure you have but ours only uses a 1% solution of > saponin, a 3% solution of calcium gluconate, and a balanced salt solution. We > get the balanced salt solution from our pharmacy. > > > Tom McNemar, HT(ASCP) > Histology Co-ordinator > Licking Memorial Health Systems > (740) 348-4163 > (740) 348-4166 > tmcne...@lmhealth.org > www.LMHealth.org > > > -Original Message- > From: histonet-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Hannen, > Valerie > Sent: Monday, October 25, 2010 11:40 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] FW: Saponin Technique > > > > > > From: Hannen, Valerie > Sent: Monday, October 25, 2010 11:39 AM > To: 'histo...@list.utsouthwestern.edu' > Subject: Saponin Technique > > > Hi folks.. > I am hoping someone out there in Histo-land can help. One of our > Pathologists is asking us to check into doing the Saponin Technique to > lyse the red cells in some of our cytology specimens. She gave a copy of > one of the pages from one of her manuals, and it shows that in order for > us to do this we would need to buy alot of reagents that we currently do > not have. Being that we would probably not be doing this technique very > often, and not sure how cost effective it will be for us to buy the > reagents, we are wondering if anyone knows of a commercial product that > we can buy that would be a cheaper way out. > > Thanks in advance!! > Valerie Hannen, Histotechnologist > Parrish Medical Center > Titusville,Florida > > > ** > "This email is intended solely for the use of the individual to > whom it is addressed and may contain information that is > privileged, confidential or otherwise exempt from disclosure > under applicable law. If the reader of this email is not the > intended recipient or the employee or agent responsible for > delivering the message to the intended recipient, you are > hereby notified that any dissemination, distribution, or > copying of this communication is strictly prohibited. If you > have received this communication in error, please immediately > delete this message. Thank you" > ** > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > This e-mail, including attachments, is intended for the sole use of the > individual and/or entity to whom it is addressed, and contains information > from Licking Memorial Health Systems which is confidential or privileged. If > you are not the intended recipient, nor authorized to receive for the > intended recipient, be aware that any disclosure, copying, distribution or > use of the contents of this e-mail and attachments is prohibited. If you have > received this in error, please advise the sender by reply e-mail and delete > the message immediately. You may also contact the LMH Process Improvement > Center at 740-348-4641. E-mail transmissions cannot be guaranteed to be > secure or error-free as information could be intercepted, corrupted, lost, > destroyed, arrive late or incomplete, or contain viruses. The sender > therefore does not accept liability for any errors or omissions in the > contents of this message, which arise as a result of e-mail transmission. > 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
RE: [Histonet] Urine Cytology
there are different slide preparation techiques used in cytology labs - ThinPrep methodology, cytocentrifugation, membrane filtration are the most often used, technique depends on pathologists preferencies and equipment and experiencies of the lab! hope this helps Irena > From: sjkit...@live.com > To: histonet@lists.utsouthwestern.edu > Date: Tue, 18 May 2010 10:52:26 -0400 > Subject: [Histonet] Urine Cytology > > > Hello, > > My company would like to start doing urine cytologies. Being a histotech i am > not 100% sure as to what i need to do this because i have to learned how to > di it yet :-) Any suggestions would be greatly appreciated! > > Thanks in Advanced > > S > > > > _ > The New Busy is not the old busy. Search, chat and e-mail from your inbox. > http://www.windowslive.com/campaign/thenewbusy?ocid=PID28326::T:WLMTAGL:ON:WL:en-US:WM_HMP:042010_3___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _ Hotmail: zanesljiva e-poštna storitev z zmogljivo Microsoftovo zaščito pred neželeno pošto https://signup.live.com/signup.aspx?id=60969___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Re: How to troubleshoot a cytology slide with dehydration problems
Dear Valerie, I can only agree that Papanicolaou stain is very tricky, having 20 yrs experiencies with it! looks very simple but you need to know some basic rules. first of all the slides should be hydrated at the beginning of staining procedure since the hematoxylin is water based stain! it seems to me thatyour slides are not enough hydrated, I never heard about sodium chloride solution as a hydrating but I guess cannot harm, however then you put slides in alcohol it's a bit confusing! and 25 seconds is a very short time for any incubation! beginning of a staining procedure depends on a fixation of the slides! in a case of a wet fixed slides you can proceed directly to staining (95% alcohol) if the slides are wet fixed and then dried you need to rehydrate them before staining and if you have spray fixed slides you need to remove the wax from slides before staining! my suggestions: 1. - leave slides in a sodium chloride longer, perhaps 2-3 minutes - alcohol 95% - alcohol 70% - water at least 3 min better 5 minutes, you'll probably need to shorten staining time in hematoxylin to 1 minute because the slides will be better hydrated and ready to accept stain. - staining time in EA solution is too short to allow proper staining and this is probably the reason to get reddish staining, you should leave the slides in EA 5 minutes since only then the light green will start to stain. hope this help! do you stain by hand or in a stainer? how often do you change stains and other solutions? you should also be very carefull to have the proper level of all solutions in a dishes! the basic rules is to keep the level of stains bellow the level of rinsing solutions. let me know about your staining results or even better to send me a picture! sometimes you can get the all redish staining also becouse of improper fixation! you can prepare your proprely fixed slide and stain it to see the results! best regards! Irena > From: vrodrigue...@gmail.com > Date: Fri, 30 Apr 2010 17:48:58 -0700 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: How to troubleshoot a cytology slide with dehydration > problems > > I forgot to include the complete protocol that my lab uses so you can help > me better: > > 1.After dehydrating in sodium chloride for 25 seconds you have to dip the > slides in alcohol 95. > 2. 10 dips in water then 1.30 minutes in hematoxylin, it used to be 1 min, > then rinse in water. > 3. 1 min in rinse solution > 4.10 dips in water. > 5. 1 min in bluing solution > 6. 10 dips in water. > 7. 10 dips in alcohol. > 8. 10 dips in alcohol. > 9. 1 min in orange solution > 10. 15 dips in 95 alcohol. > 11. 15 dips in 95 alcohol. > 12. 3.15 mins in EA solution > 13. The slides are then dipped in 3 containers of 95 alcohol (20 dips each) > 14. Slides are dipped in 3 containers of 99 alcohol (15 dips in each) > 15. Then they are dipped in Isoxylene which is a combination of xylene and > 99 alcohol (15 dips) > 16. And lastly Xylene 10 dips > > > When the slides do not absorb the h20 saline seems that the slides absorb > more EA than hematoxylin. I don't know why. > > On Fri, Apr 30, 2010 at 5:34 PM, Valerie R. wrote: > > > I am an HTL but I have to stain pap smears. In the Papanicalou procedure > > for fine needle aspirations I have to stain the slides in sodium chloride > > first for 25 seconds in total (15 seconds in one container and 10 in another > > container). This for me is the trickiest part when staining cytology because > > if slides are not well dehydrated in saline then the cells are not going to > > absorb the stains well, and the result will be a pinkish slide, when it is > > suppose to look blue-greenish. > > > > I had this problem today where I stained an entire rack of slides, and all > > the slides turned blue (which means they stained correctly) except 4 slides > > which turned out pinkish (which is a sign that they did not dehydrated > > properly in sodium chloride (h20 saline). > > > > Is there a solution to this issue. Can these slides be re-stained? > > > > I know this newsgroup is for histology only but I would like to know if > > histotech and cytotechs have encountered this issue in their labs and how > > they ended up solving the problem. > > > > Thanks in advance > > > > > > > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _ Hotmail: zmogljiva brezplačna e-poštna storitev z Microsoftovo zaščito https://signup.live.com/signup.aspx?id=60969___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
