[Histonet] Tissue-Tek cover slipper question
Good morning. I work part-time at a small lab and they have a Sakura Feintek Tissue-Tek SCA tape coverslipper, model 4764. I have not worked with any tape coverslippers before and am having problems with areas of the tissue drying out. The solvent bottle is full of Xylene. However, there is a crack in the lid of the bottle. After removing the stained slides from the autostainer and loading the basket(s) to place on the cover slipper, I re-dip the slides in Xylene. The room where this equipment is housed, does get warm. I was wondering if the tape can become old brittle, leading to it not sealing well, or if not enough Xylene is being deposited on each slide or a combination of both? I could not find the manual on the unit for trouble shooting ideas. Any help would be greatly appreciated. Thank you Candy Candy Bales, HT Chief Histologist The University of Texas Dental Branch at Houston Diagnostic Sciences-Oral Pathology 6516 M.D. Anderson Blvd. # 3.093 Houston, TX 77030 713.500.4411 office 713.500.4416 fax ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] remove me from list
Please remove this email from the list. sad...@hcmhcares.org -- *Sharon Adams* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Tissue-Tek cover slipper question
Are you letting the tape sit out (cure) for awhile befoe you use it? We leave ours out overnight? Lisa -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bales, Candy A Sent: Tuesday, April 12, 2011 8:19 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue-Tek cover slipper question Good morning. I work part-time at a small lab and they have a Sakura Feintek Tissue-Tek SCA tape coverslipper, model 4764. I have not worked with any tape coverslippers before and am having problems with areas of the tissue drying out. The solvent bottle is full of Xylene. However, there is a crack in the lid of the bottle. After removing the stained slides from the autostainer and loading the basket(s) to place on the cover slipper, I re-dip the slides in Xylene. The room where this equipment is housed, does get warm. I was wondering if the tape can become old brittle, leading to it not sealing well, or if not enough Xylene is being deposited on each slide or a combination of both? I could not find the manual on the unit for trouble shooting ideas. Any help would be greatly appreciated. Thank you Candy Candy Bales, HT Chief Histologist The University of Texas Dental Branch at Houston Diagnostic Sciences-Oral Pathology 6516 M.D. Anderson Blvd. # 3.093 Houston, TX 77030 713.500.4411 office 713.500.4416 fax ___ 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] Her2 FISH controls
Hello, I see that the ASCO/CAP Guidelines recommend using amplified, cutoff, and normal controls for Her2 FISH testing. We currently use Pathvysion and purchase normal and cutoff controls from Abbott. They do not offer a product of amplified controls. Are you all running this third control? Are you using in-house tissue? Have you found a vendor who sells them? Thanx Melissa Kuhnla Lead Medical Technologist for IHC and FISH Catholic Health Services of Long Island Regional Laboratory Services The information in this e-mail, and any attachments therein, is confidential and for use by the intended addressee only. If this message is received by you in error please do not disseminate or read further. Please reply to the sender that you have received the message in error, then delete the message. Although Catholic Health Services of Long Island attempts to sweep e-mail and attachments for viruses, it does not guarantee that either are virus-free and accepts no liability for any damage sustained as a result of viruses. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Tiisue tek coverslipper
Thank you all for your suggestions. I will observe the drip rate and increase it as well as replacing the cracked lid on the solvent bottle. I did leave the tape out for a couple of days before placing it on the machine as a friend who has a different model suggested I do so. Thank you again Histonetters candy Candy Bales, HT Chief Histologist The University of Texas Dental Branch at Houston Diagnostic Sciences-Oral Pathology 6516 M.D. Anderson Blvd. # 3.093 Houston, TX 77030 713.500.4411 office 713.500.4416 fax ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Counterstain for dual chromogenic
Good morning, I have been attempting to do some dual chromogenic staining. I am using a mouse and a rabbit antibody. For the mouse antibody I am using an HRP-mouse secondary followed by AEC for visualization. For the rabbit antibody I am using a biotinylated rabbit secondary, followed by ABC-AP and vector blue. My problem is what to use as a counterstain. From everything I have read so far there isn't one that would work without having some problems. I was thinking of trying a very light Hematoxylin and see if it doesn't disrupt the vector blue visualization too much. Does anyone else have any suggestions? Thanks, Eva ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] microwave processors
Has anyone used the LabPulse microwave processor from EBS? We need a new microwave for staining and retrieval and thought maybe we could get one that will do processing also. I've looked at the Shurwave from Fisher, and the Mars from Hacker. Are there any other companies that have small microwave processors? Margaret Perry HT(ASCP) Dept of Veterinary and Biomedical services Box 2175 South Dakota State University Brookings SD 57007 605-688-5638 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: NY cert requirements
That is a huge question. Here is a good place to start NYS Clinical Laboratory TechnologySep 9, 2009 ... Clinical Laboratory Technician. License Requirements ... www.op.nysed.gov Loralee McMahon, HTL (ASCP) Immunohistochemistry Supervisor Strong Memorial Hospital Department of Surgical Pathology (585) 275-7210 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Pitts, Jaclyn S. (Jackie), HT(ASCP) [pitts.jac...@mayo.edu] Sent: Monday, April 11, 2011 9:32 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] NY cert requirements Hello, My colleague and I have been wondering what the certification requirements are in New York and what other kind of guidelines that have to be followed for the state as well in the lab. Jaclyn Pitts, HT(ASCP) Histotechnician E-mail: pitts.jac...@mayo.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] Counterstain for dual chromogenic
You could do that or you could use DAB for one antibody (brownish color), followed by DAB + Nickel for the other (bluish color) and they are both permanent and alcohol resistant. René J. --- On Tue, 4/12/11, Eva Permaul e...@georgetown.edu wrote: From: Eva Permaul e...@georgetown.edu Subject: [Histonet] Counterstain for dual chromogenic To: histonet@lists.utsouthwestern.edu Date: Tuesday, April 12, 2011, 11:37 AM Good morning, I have been attempting to do some dual chromogenic staining. I am using a mouse and a rabbit antibody. For the mouse antibody I am using an HRP-mouse secondary followed by AEC for visualization. For the rabbit antibody I am using a biotinylated rabbit secondary, followed by ABC-AP and vector blue. My problem is what to use as a counterstain. From everything I have read so far there isn't one that would work without having some problems. I was thinking of trying a very light Hematoxylin and see if it doesn't disrupt the vector blue visualization too much. Does anyone else have any suggestions? Thanks, Eva ___ 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] microwave processors
Milestone manufactures very good and versatile MW that process and stain. René J. --- On Tue, 4/12/11, Perry, Margaret margaret.pe...@sdstate.edu wrote: From: Perry, Margaret margaret.pe...@sdstate.edu Subject: [Histonet] microwave processors To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Date: Tuesday, April 12, 2011, 12:08 PM Has anyone used the LabPulse microwave processor from EBS? We need a new microwave for staining and retrieval and thought maybe we could get one that will do processing also. I've looked at the Shurwave from Fisher, and the Mars from Hacker. Are there any other companies that have small microwave processors? Margaret Perry HT(ASCP) Dept of Veterinary and Biomedical services Box 2175 South Dakota State University Brookings SD 57007 605-688-5638 ___ 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
AW: [Histonet] Her2 FISH controls
We use inhouse tissue and made really tiny pieces to place them near the specimen slide. So we bring it beneath one coverslip. Bye Gudrun -Ursprüngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Kuhnla, Melissa Gesendet: Dienstag, 12. April 2011 15:36 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Her2 FISH controls Hello, I see that the ASCO/CAP Guidelines recommend using amplified, cutoff, and normal controls for Her2 FISH testing. We currently use Pathvysion and purchase normal and cutoff controls from Abbott. They do not offer a product of amplified controls. Are you all running this third control? Are you using in-house tissue? Have you found a vendor who sells them? Thanx Melissa Kuhnla Lead Medical Technologist for IHC and FISH Catholic Health Services of Long Island Regional Laboratory Services The information in this e-mail, and any attachments therein, is confidential and for use by the intended addressee only. If this message is received by you in error please do not disseminate or read further. Please reply to the sender that you have received the message in error, then delete the message. Although Catholic Health Services of Long Island attempts to sweep e-mail and attachments for viruses, it does not guarantee that either are virus-free and accepts no liability for any damage sustained as a result of viruses. 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] Benchmark Ultra question - follow up
Today I made a further experiment. I did two titration runs. On one slide I added the working solution by pipette on the other slide I added it by pushing the filled PrepKit. Both stainings came out beautiful. - There has to be something wrong with the dispension-amount. Ventana is informed, I hope they find the culprit. Bye Gudrun -Ursprüngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Gudrun Lang Gesendet: Montag, 11. April 2011 18:48 An: 'Angela Bitting' Cc: histonet@lists.utsouthwestern.edu Betreff: AW: [Histonet] Benchmark Ultra question Yes, that could be true. Some of the PrepKits go harder than others. Perhaps the pressure of the hammer is not big enough. Sometimes the nozzles are not filled equally. (We press always reagens in the nozzle when we prepare the run.) With the Ventana-System there is an additional dilution, because the working-solution is added to a reaction-buffer film under the LCS. Perhaps it makes a difference if you dispense the solution with the pipett-tip under the LCS or if the drop falls on the surface of the LCS. That could be a matter of the LCS-quality, or buffer-quantity , or . I think I'll become mad! Our pathologists have an uncertain unhappiness with the overall staining results, but most of the antibodies work well. Perhaps the majority isn't very sensitiv to small changes in the system and the quality-difference isn't big enough, but some are easier to kill. Regards, Gudrun _ Von: Angela Bitting [mailto:akbitt...@geisinger.edu] Gesendet: Montag, 11. April 2011 18:06 An: gu.l...@gmx.at Betreff: Re: [Histonet] Benchmark Ultra question When I was trained to do titration runs on the BenchmarkXT and Ultras, I was told to titrate 100ul of antibody. I have been suspecting for some time now that the dispensers DON'T dispense a full 100ul. I haven't taken the time to prove it, but your question may have motivated me. That could explain the weak staining. Gudrun Lang gu.l...@gmx.at 4/11/2011 11:48 AM Hi! I have some issues with our new Ultra. Since a few days antibodies, that work usually well, show up very faint. For example the TTF1 (Novocastra). I ordered a new bottle, made a titration and found, that the old titer 1:50 was again well enough. I filled the old PrepKit - and the result was very weak. Then I thought, the PrepKit itself is the culprit. I changed the PrepKit - and again the result was very weak. At that time the working-solution was only three days old. Has anyone an explanation, why the titration works well and the automated dispension doesn't? Regards Gudrun Lang ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _ IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. 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] Milestone MW
Margret, Which microwave would that be THANK YOU, PATTI RUBEN-NELSON H.T.(ASCP) LABORATORY CONSULTANT SUPERVISOR/DESERT GASTROENTEROLOGY CONSULTANTS 35-900 Bob Hope Drive Suite 275 Rancho Mirage, Ca. 92270 cell (909) 841-9761 / wk (760) 321-2500 nelsonr...@verizon.net ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Immunofluorescence and FISH Combined stating
Where can I find a procedure for dual staining of blood and bone marrow smears using Vector anti-kappa and anti-lambda tagged with Coumarine followed by FISH hybridization. I am able to see what I think are plasma cells by their fluorescent blue cytoplasm and on the same slide (changing filters) I can see the red and green signal patterns. From: : di...@bioview.co.il ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Von kossa stain
What is everyone using for their light when developing the silver in the VonKossa stain when you have no sunlight to use? We used to use a 60 watt lamp, but haven't done one for years and am bringing this stain back to our repetiore due to pathologist request. Thanks much! Dorothy Webb, HT (ASCP) Regions Histology Technical Supervisor 651-254-2962 This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Von kossa stain
Funny you should ask... just last week I had two requests for a Von Kossa. That's when I found out that, despite the fact that we just moved into a brand new building with all the bells and whistles, there was not one single UV light in any of the many hoods that had been installed. Heaven forbid I could even find an incandescent bulb (we are a green building). So, I prepped my slides, put them in a clear glass Coplin jar and parked the jar on the hood of my car for an hour. Works like a charm. This all depends, naturally, on (1) if you even HAVE sunshine where you live; (2) how far a walk it is to the sunshine, and (3) whether you have a car hood on which to park the Coplin jar. Needless to say, out of the line of sight of curious onlookers with sticky fingers and no business wondering what that glass jar is... Nonetheless, it works just fine! May the Force be with you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Histonet Digest, Vol 89, Issue 12
Date: Tue, 12 Apr 2011 11:37:17 -0400 From: Eva Permaul e...@georgetown.edu Subject: [Histonet] Counterstain for dual chromogenic To: histonet@lists.utsouthwestern.edu Message-ID: 4da471ad.3040...@georgetown.edu Content-Type: text/plain; charset=ISO-8859-1; format=flowed Eva I have been doing some double labelling using Vulcan Fast Red from Biocare and Vector Blue Alkaline Phosphatase Substrate Kit III from Vector Labs. For counter stain I use Vector Methyl Green and the combination works great and looks quite nice. Mark Good morning, I have been attempting to do some dual chromogenic staining. I am using a mouse and a rabbit antibody. For the mouse antibody I am using an HRP-mouse secondary followed by AEC for visualization. For the rabbit antibody I am using a biotinylated rabbit secondary, followed by ABC-AP and vector blue. My problem is what to use as a counterstain. From everything I have read so far there isn't one that would work without having some problems. I was thinking of trying a very light Hematoxylin and see if it doesn't disrupt the vector blue visualization too much. Does anyone else have any suggestions? Thanks, Eva ***CONFIDENTIALITY NOTICE*** This electronic message and any attachments are intended only for the use of the addressee and may contain information that is privileged and confidential. Any dissemination, distribution or copying of this communication by unauthorized individuals is strictly prohibited. If you have received this communication in error, please notify the sender immediately by reply e-mail and delete the original and all copies from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
AW: [Histonet] Von kossa stain
WE use just a simple desk-lamp and put the coplin jar directly under the bulb. And the desk-lamp is placed in the window. Bye Gudrun -Ursprüngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Webb, Dorothy L Gesendet: Dienstag, 12. April 2011 19:35 An: 'histonet@lists.utsouthwestern.edu' Betreff: [Histonet] Von kossa stain What is everyone using for their light when developing the silver in the VonKossa stain when you have no sunlight to use? We used to use a 60 watt lamp, but haven't done one for years and am bringing this stain back to our repetiore due to pathologist request. Thanks much! Dorothy Webb, HT (ASCP) Regions Histology Technical Supervisor 651-254-2962 This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ 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: AW: [Histonet] Von kossa stain
We have intense sun here but my windows face the wrong way! At least I have windows. I just use a light bulb from the lamp we have over the coverslipping station. Works great. Andi On Apr 12, 2011, at 11:22 AM, Gudrun Lang wrote: WE use just a simple desk-lamp and put the coplin jar directly under the bulb. And the desk-lamp is placed in the window. Bye Gudrun -Ursprüngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Webb, Dorothy L Gesendet: Dienstag, 12. April 2011 19:35 An: 'histonet@lists.utsouthwestern.edu' Betreff: [Histonet] Von kossa stain What is everyone using for their light when developing the silver in the VonKossa stain when you have no sunlight to use? We used to use a 60 watt lamp, but haven't done one for years and am bringing this stain back to our repetiore due to pathologist request. Thanks much! Dorothy Webb, HT (ASCP) Regions Histology Technical Supervisor 651-254-2962 This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ 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
[Histonet] Staining using a Plastic Sytem
Hi, Does anyone have experience embedding and staining in plastic? We have rabbit rotator cuff tendons that are being embedded in plastic. I am trying to figure out what the process is and which stains to use in order to visualize (and quantify) blood vessels and collagen deposition at the Bone (Humeral Head) - Tendon (Infraspinatus) interface. Thanks, Mahesh ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Biopsy program for Sakura VIP 5/6 Processor
Our lab is currently looking for a way to shorten our Biopsy processing program without compromising the patient specimen. We do have an issue with our GI's being very dry, which causes us to have to soak between each level taken and also causes a lot of chatter. Also we have a goal to do a run during the day to improve turn around time. Here is what our current protocol is Formalin1 min 37degrees Formalin15 mins37degrees 70 14 mins40 degrees 95 14 mins40 degrees 95 9 mins 40 degrees 100 9 mins 40 degrees 100 7 mins 40 degrees 100 4 mins 40 degrees Xylene 23 minsno heat Xylene 15 minsno heat Paraffin 20 mins 60 degrees Paraffin 18 mins 60 degrees Paraffin 10 mins 60 degrees Paraffin 0 mins 60 degrees All the steps are set on a fast mix setting. All of our biopsy specimens are put into sponges. Any feedback would be greatly appreciated. Andria B Evans HTL(ASCP)CM Lancaster General Hospital 555 North Duke Street Lancaster, PA 17604 (717)544-5511 ext: 77329 aeva...@lgheath.orgmailto:aeva...@lgheath.org This email was sent securely from the LGHealth Email Service Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact 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
Re: [Histonet] Milestone MW
Model KOS by Milestone is capable of tissue processing, decalcification, special stains, fixation, gross hardening, and antigen retrieval. René J. --- On Tue, 4/12/11, SHANE NELSON nelsonr...@verizon.net wrote: From: SHANE NELSON nelsonr...@verizon.net Subject: [Histonet] Milestone MW To: Histonet@lists.utsouthwestern.edu Date: Tuesday, April 12, 2011, 12:30 PM Margret, Which microwave would that be THANK YOU, PATTI RUBEN-NELSON H.T.(ASCP) LABORATORY CONSULTANT SUPERVISOR/DESERT GASTROENTEROLOGY CONSULTANTS 35-900 Bob Hope Drive Suite 275 Rancho Mirage, Ca. 92270 cell (909) 841-9761 / wk (760) 321-2500 nelsonr...@verizon.net ___ 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] Von kossa stain
You can use any strong intensity microscope light. The stronger the intensity of the light the better, but the reduction has a cumulative effect so even not very strong intensity bulbs can be effective. René J. --- On Tue, 4/12/11, Webb, Dorothy L dorothy.l.w...@healthpartners.com wrote: From: Webb, Dorothy L dorothy.l.w...@healthpartners.com Subject: [Histonet] Von kossa stain To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu Date: Tuesday, April 12, 2011, 1:35 PM What is everyone using for their light when developing the silver in the VonKossa stain when you have no sunlight to use? We used to use a 60 watt lamp, but haven't done one for years and am bringing this stain back to our repetiore due to pathologist request. Thanks much! Dorothy Webb, HT (ASCP) Regions Histology Technical Supervisor 651-254-2962 This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ 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] Von kossa stain
I purchase a kit from Dorn and Hart Microedge and use sodium carbonate-formaldehyde solution to chemically develop. Stain in silver nitrate for 5 min in the dark, 3 fresh DI rinses for 1 min each in the dark, then sodium carbonate-formaldehyde solution for 2 min in the dark, 2 fresh DI rinses (normal light) for 1 min each, 30 seconds in Farmer's Diminisher (Sodium thiosulfate-potassium ferricyanide soluiton to stop reaction) and a running tap water rinse for 10 min. Jack From: dorothy.l.w...@healthpartners.com To: histonet@lists.utsouthwestern.edu Date: Tue, 12 Apr 2011 12:35:18 -0500 Subject: [Histonet] Von kossa stain What is everyone using for their light when developing the silver in the VonKossa stain when you have no sunlight to use? We used to use a 60 watt lamp, but haven't done one for years and am bringing this stain back to our repetiore due to pathologist request. Thanks much! Dorothy Webb, HT (ASCP) Regions Histology Technical Supervisor 651-254-2962 This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0 ___ 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] Counterstain for dual chromogenic
Try ethyl green: 1. 0.1N Acetic Acid Add 6ml of glacial acetic acid to 1000 ml of distilled water. 2. 0.1N Sodium Acetate Add 4.102 g of sodium acetate to 500 ml of distilled water 3. pH 4.2 buffer Add 755 ml of 0.1N acetic acid to 264 ml of 0.1N Sodium Acetate and adjust the pH to 4.2 with 1M NaOH 4. Ethyl Green Solution To 100ml of buffer add 2g Ethyl Green (CI 42590) Can be stored at room temp. If staining turns bluish than the solution has started to deteriorate. Stain for 5 minutes. Do not rinse in water for too long, it tends to extract the green staining Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Eva Permaul Sent: Wednesday, 13 April 2011 1:37 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Counterstain for dual chromogenic Good morning, I have been attempting to do some dual chromogenic staining. I am using a mouse and a rabbit antibody. For the mouse antibody I am using an HRP-mouse secondary followed by AEC for visualization. For the rabbit antibody I am using a biotinylated rabbit secondary, followed by ABC-AP and vector blue. My problem is what to use as a counterstain. From everything I have read so far there isn't one that would work without having some problems. I was thinking of trying a very light Hematoxylin and see if it doesn't disrupt the vector blue visualization too much. Does anyone else have any suggestions? Thanks, Eva ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Von kossa stain
Interesting, Meloan Puchtler (1985) have drawn our attention to Von Kossa's original work on this technique. He regarded only the yellow colouration of calcium deposits during early stages of the reaction as diagnostic for calcium phosphate and credited the blackening to organic matter. Further studies showed that bright light only causes the irreversible blackening of organic matter that masks the yellow silver phosphate. When the reaction is performed in subdued light, yellow to yellowish brown silver phosphate is visualised selectively. Silver carbonate dissolves in sodium thiosulphate and cannot be demonstrated with von Kossa's technique (Meloan Puchtler 1985 J Histotechnol 8(1):11-13.). Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Breeden, Sara Sent: Wednesday, 13 April 2011 3:53 AM To: Webb, Dorothy L; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Von kossa stain Funny you should ask... just last week I had two requests for a Von Kossa. That's when I found out that, despite the fact that we just moved into a brand new building with all the bells and whistles, there was not one single UV light in any of the many hoods that had been installed. Heaven forbid I could even find an incandescent bulb (we are a green building). So, I prepped my slides, put them in a clear glass Coplin jar and parked the jar on the hood of my car for an hour. Works like a charm. This all depends, naturally, on (1) if you even HAVE sunshine where you live; (2) how far a walk it is to the sunshine, and (3) whether you have a car hood on which to park the Coplin jar. Needless to say, out of the line of sight of curious onlookers with sticky fingers and no business wondering what that glass jar is... Nonetheless, it works just fine! May the Force be with you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Biopsy program for Sakura VIP 5/6 Processor
Increase the fixation time and make sure the air is out of the sponges (will stop formalin from getting to the tissue- a quick dunk of the cassette (with sponge and tissue) in alcohol and back into formalin will do the trick). Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Evans, Andria B Sent: Wednesday, 13 April 2011 4:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Biopsy program for Sakura VIP 5/6 Processor Our lab is currently looking for a way to shorten our Biopsy processing program without compromising the patient specimen. We do have an issue with our GI's being very dry, which causes us to have to soak between each level taken and also causes a lot of chatter. Also we have a goal to do a run during the day to improve turn around time. Here is what our current protocol is Formalin1 min 37degrees Formalin15 mins37degrees 70 14 mins40 degrees 95 14 mins40 degrees 95 9 mins 40 degrees 100 9 mins 40 degrees 100 7 mins 40 degrees 100 4 mins 40 degrees Xylene 23 minsno heat Xylene 15 minsno heat Paraffin 20 mins 60 degrees Paraffin 18 mins 60 degrees Paraffin 10 mins 60 degrees Paraffin 0 mins 60 degrees All the steps are set on a fast mix setting. All of our biopsy specimens are put into sponges. Any feedback would be greatly appreciated. Andria B Evans HTL(ASCP)CM Lancaster General Hospital 555 North Duke Street Lancaster, PA 17604 (717)544-5511 ext: 77329 aeva...@lgheath.orgmailto:aeva...@lgheath.org This email was sent securely from the LGHealth Email Service Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact 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 * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] RE: Biopsy program for Sakura VIP 5/6 Processor
I would consider replacing the sponges. Tony is correct that you need to make sure the air is removed from the sponge to facilitate exchange, but with shortened processing times you will undoubtedly cause carry over from solution to solution. Yuo may want to have a small container of alcoholic formalin with sponges sitting on the gross table. Try using a nylon tissue bag to funnel filter your small biopsies. all your biopsy sample sizes can be placed into the bag and very tiny samples are retained. Last suggestion is to consider a folded filter paper method (we name it origami, I can provide directly if you want) that uses a quick four fold process w/ forceps at the gross bench to create a pocket for the tiny samples, has a single layer between the tissue/solution and is easy to open at embedding (without popping of tissue samples). Avoind the unnecessary carryover of the sponge if you can. William DeSalvo, B.S., HTL(ASCP) From: antho...@chw.edu.au To: aeva...@lghealth.org; histonet@lists.utsouthwestern.edu Date: Tue, 12 Apr 2011 23:30:30 + CC: Subject: [Histonet] RE: Biopsy program for Sakura VIP 5/6 Processor Increase the fixation time and make sure the air is out of the sponges (will stop formalin from getting to the tissue- a quick dunk of the cassette (with sponge and tissue) in alcohol and back into formalin will do the trick). Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Evans, Andria B Sent: Wednesday, 13 April 2011 4:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Biopsy program for Sakura VIP 5/6 Processor Our lab is currently looking for a way to shorten our Biopsy processing program without compromising the patient specimen. We do have an issue with our GI's being very dry, which causes us to have to soak between each level taken and also causes a lot of chatter. Also we have a goal to do a run during the day to improve turn around time. Here is what our current protocol is Formalin1 min 37degrees Formalin15 mins37degrees 70 14 mins40 degrees 95 14 mins40 degrees 95 9 mins 40 degrees 100 9 mins 40 degrees 100 7 mins 40 degrees 100 4 mins 40 degrees Xylene 23 minsno heat Xylene 15 minsno heat Paraffin 20 mins 60 degrees Paraffin 18 mins 60 degrees Paraffin 10 mins 60 degrees Paraffin 0 mins 60 degrees All the steps are set on a fast mix setting. All of our biopsy specimens are put into sponges. Any feedback would be greatly appreciated. Andria B Evans HTL(ASCP)CM Lancaster General Hospital 555 North Duke Street Lancaster, PA 17604 (717)544-5511 ext: 77329 aeva...@lgheath.orgmailto:aeva...@lgheath.org This email was sent securely from the LGHealth Email Service Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact 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 * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list
RE: [Histonet] RE: Biopsy program for Sakura VIP 5/6 Processor
I agree Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA From: WILLIAM DESALVO [mailto:wdesalvo@hotmail.com] Sent: Wednesday, 13 April 2011 9:59 AM To: Tony Henwood; aeva...@lghealth.org; histonet Subject: RE: [Histonet] RE: Biopsy program for Sakura VIP 5/6 Processor I would consider replacing the sponges. Tony is correct that you need to make sure the air is removed from the sponge to facilitate exchange, but with shortened processing times you will undoubtedly cause carry over from solution to solution. Yuo may want to have a small container of alcoholic formalin with sponges sitting on the gross table. Try using a nylon tissue bag to funnel filter your small biopsies. all your biopsy sample sizes can be placed into the bag and very tiny samples are retained. Last suggestion is to consider a folded filter paper method (we name it origami, I can provide directly if you want) that uses a quick four fold process w/ forceps at the gross bench to create a pocket for the tiny samples, has a single layer between the tissue/solution and is easy to open at embedding (without popping of tissue samples). Avoind the unnecessary carryover of the sponge if you can. William DeSalvo, B.S., HTL(ASCP) From: antho...@chw.edu.aumailto:antho...@chw.edu.au To: aeva...@lghealth.orgmailto:aeva...@lghealth.org; histonet@lists.utsouthwestern.edumailto:histonet@lists.utsouthwestern.edu Date: Tue, 12 Apr 2011 23:30:30 + CC: Subject: [Histonet] RE: Biopsy program for Sakura VIP 5/6 Processor Increase the fixation time and make sure the air is out of the sponges (will stop formalin from getting to the tissue- a quick dunk of the cassette (with sponge and tissue) in alcohol and back into formalin will do the trick). Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edumailto:histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu]mailto:[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Evans, Andria B Sent: Wednesday, 13 April 2011 4:57 AM To: histonet@lists.utsouthwestern.edumailto:histonet@lists.utsouthwestern.edu Subject: [Histonet] Biopsy program for Sakura VIP 5/6 Processor Our lab is currently looking for a way to shorten our Biopsy processing program without compromising the patient specimen. We do have an issue with our GI's being very dry, which causes us to have to soak between each level taken and also causes a lot of chatter. Also we have a goal to do a run during the day to improve turn around time. Here is what our current protocol is Formalin 1 min 37degrees Formalin 15 mins 37degrees 70 14 mins 40 degrees 95 14 mins 40 degrees 95 9 mins 40 degrees 100 9 mins 40 degrees 100 7 mins 40 degrees 100 4 mins 40 degrees Xylene 23 mins no heat Xylene 15 mins no heat Paraffin 20 mins 60 degrees Paraffin 18 mins 60 degrees Paraffin 10 mins 60 degrees Paraffin 0 mins 60 degrees All the steps are set on a fast mix setting. All of our biopsy specimens are put into sponges. Any feedback would be greatly appreciated. Andria B Evans HTL(ASCP)CM Lancaster General Hospital 555 North Duke Street Lancaster, PA 17604 (717)544-5511 ext: 77329 aeva...@lgheath.orgmailto:aeva...@lgheath.orgmailto:aeva...@lgheath.org%3cmailto:aeva...@lgheath.org This email was sent securely from the LGHealth Email Service Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edumailto:Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at
[Histonet] Update regarding question on Plastic Embedding
Looking for a protocol to visualize vascularization and collagen deposition at the bone-tendon interface of a rabbit rotator cuff embedded in a plastic system. Sections will be rather thick (~50um) b/c they are being made through a titanium anchor. Using MMA with a cold-curing resin, Technovit 9100. Thanks in advance! - Mahesh ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] FW: 1 HE slide vs. 2
From: eugenia90...@hotmail.com To: histonet@lists.utsouthwestern.edu Subject: 1 HE slide vs. 2 Date: Mon, 11 Apr 2011 13:06:59 -0400 Good afternoon everyone, Does anyone know of an article or statistics discussing the impact (medical diagnosing) of cutting 2 HE slides per block verses 1 for all routine work? Genia ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
