[Histonet] brain sections falling off slides during IHC
A few years ago we also had this problem. The brains were terrible. We found that changing our processing chemicals, especially the wax, more frequently got rid of most of our problems. We use Fisher plus slides. There is a difference in plus slides. I recommend you contact several distributors and get samples and pick the one that works best for you. After cutting we have a fan that air dries the slides until the sections look dry about 10- 15 min. The slides are dried overnight on top of our embedder so that they are heated slightly. The next day they are dried for 1 hour at 60° C. We developed this method for cattle ear notches and about the only reason tissue comes off the slide now is because the tissue is necrotic or not fixed properly. Margaret Perry HT (ASCP) IHC Lab Manager Veterinary Science Animal Disease Research and Diagnostic Lab South Dakota State University Box 2175 North Campus Drive Brookings SD 57007 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Subject: [Histonet] brain sections falling off slides during IHC
What type of plus slides are you using? There are a few companies out there selling a "super" adhesive slide for just such applications. We use the Bond380 slide from TruScientific for all of our brain, autopsy and fatty tissue when performing IHC. We rarely have any compromised tissue and that is using the Dako platform and the PT Module for retrieval. I know there are other companies that carry similar slides, but we found the best pricing on these. You can reach them at 877-677-7523. You might also try skipping the oven and air drying your sections overnight follwed by a run over the heat plate before starting the staining process. Jennifer Bull jennifer.b...@nwpathology.com -Original Message- From: histonet-bounces <@t> lists.utsouthwestern.edu<http://lists.utsouthwestern.edu/mailman/listinfo/histonet> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu<http://lists.utsouthwestern.edu/mailman/listinfo/histonet>] On Behalf Of Thurby, Christina Sent: Wednesday, February 11, 2009 7:38 AM To: histonet <@t> lists.utsouthwestern.edu<http://lists.utsouthwestern.edu/mailman/listinfo/histonet> Subject: [Histonet] brain sections falling off slides during IHC Hello all, Can any of you all with experience immunostaining brain sections please help. My sections are almost completely washing off the slides. I am using 10% NBF fixed canine brain sections. Embedded in paraffin and sectioned at 4 microns on plus slides with distilled water in the water bath. The slides are dried at 65 C for 15 minutes and allowed to stand overnight. I am using a Dako Autostainer for staining. There is no antigen retrieval step. My staining procedure is approximately 3.5 hours long. At the completion of the procedure, the tissue on my slides is 50-80% washed off. Any thoughts?? Christina Thurby ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] brain sections falling off slides during IHC
Hi, my experience is to use lower concentration gel-coated slides or pre-coated slide purchased from company...dont use home made one if you want to improve. second, the heating procedure shortly after mounting the frozen sections onto slides does not help a lot. My way: mounting sections from cryostat onto slide -> air dry -> heat plate 40-50 C or not for 10 min until the slides are a bit hot -> air dry in fume hood overnight (for small section such as Olfactory bulb, coronal brain section of rat takes 1-2 days - much better) ->you can stock all sections in -20 C freezer ->each time before IHC, heat the section at 60 C for 10 min (dry up after taking out from freezer). I boil the slide in 95 C citrate buffer for 30 min, <1 sections from 100 falls off. 2009-02-12 TF 发件人： Thurby, Christina 发送时间： 2009-02-11 23:43:16 收件人： histonet@lists.utsouthwestern.edu 抄送： 主题： [Histonet] brain sections falling off slides during IHC Hello all, Can any of you all with experience immunostaining brain sections please help. My sections are almost completely washing off the slides. I am using 10% NBF fixed canine brain sections. Embedded in paraffin and sectioned at 4 microns on plus slides with distilled water in the water bath. The slides are dried at 65 C for 15 minutes and allowed to stand overnight. I am using a Dako Autostainer for staining. There is no antigen retrieval step. My staining procedure is approximately 3.5 hours long. At the completion of the procedure, the tissue on my slides is 50-80% washed off. Any thoughts?? Christina Thurby ___ 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] brain sections falling off slides during IHC
Hello all, Can any of you all with experience immunostaining brain sections please help. My sections are almost completely washing off the slides. I am using 10% NBF fixed canine brain sections. Embedded in paraffin and sectioned at 4 microns on plus slides with distilled water in the water bath. The slides are dried at 65 C for 15 minutes and allowed to stand overnight. I am using a Dako Autostainer for staining. There is no antigen retrieval step. My staining procedure is approximately 3.5 hours long. At the completion of the procedure, the tissue on my slides is 50-80% washed off. Any thoughts?? Christina Thurby ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
