RE: [Histonet] Losing tissue on IHC slides
Good Morning Another thing to try is different slides. We have had issues with bad batches/lots of slides - even the Fisher Superfrost plus. Especially if your purchasing department buys in quantity and then stores the slides in hot warehouses. This seems to negate the charge on the slides - but in an inconsistent way. You can have bad slides mixed with good ones in the same box. Even if you don't want to change vendors, trying a different kind of charged slide would eliminate bad slides as part of the problem. And just an FYI- we do our control slides the same way you do, (and use tap water) so I don't think that is your problem. We are using the "cheaper" Fisher Superfrost plus slides and they seem to work as well as the full priced ones. We cut our immuno sections at 3um. I know how frustrating this can be - I feel your pain! Good luck! Sue Sue Hunter, Supervisor Advanced Diagnostics Beaumont Health System Royal Oak MI 248-898-5146 shun...@beaumont.edu -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Deloris Carter Sent: Friday, July 26, 2013 5:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Losing tissue on IHC slides Hi all, I'm looking for some troubleshooting help. In the last couple of months we've been having an increasing trend in losing tissue from our IHC slides. We use a Ventana Benchmark XT. PM is done on it quarterly. The problem seems to be with the patient tissue, not the control tissue. We use Fisherbrand Superfrost Plus slides, picking up the control by dipping the top end of the slide to avoid "double dipping". We use tap water in the waterbaths which I know could cause an issue, but that's the way it's always been done here, so there's been no change to that part of the procedure. (We did try using DI water, but it didn't make any difference. Tissue still fell off of slides.) We do use DI water to mix the bulk reagents.The control slides may sometimes have been cut for a longer than recommended time, but that part of the procedure hasn't changed either, same process as always. If we run separate slides for control and patient tissue, we get much better results, but still have some tissue loss. I can't say it's all of our tests, but it is a steadily worsening problem. I'm going through tons of reagent, and I can't seem to nail down the problem. Ventana rep said to try DI water, don't double dip, try adhesive slides (which didn't really make any difference that we could tell). I would appreciate any suggestions Deloris Carter Shawnee Mission Medical Center ___ 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] Losing tissue on IHC slides
I have a couple of things. First off, are the sections coming off the slides entirely, or does it have a "chewed up" appearance? If the sections are coming completely off, I would agree with the others that water is the issue. Also, make sure you are cutting no thicker than 4 microns. We had repeated a test twice due to sections falling off, and the tech who cut them has a tendency to cut on 5 microns. I cut them the third time at 4 microns, and let the sections really flatten out on the water bath for a minute. Then baked them well, and third time was the charm. The sections remained on. As far as the "double dipping" of precut control slides go, I spoke directly with a representative from Erie (the makers of all superfrost plus slides) and was told you could dip those slides a hundred times in water, it will not affect the charge on the slides. We've never worried about double dipping and it has never caused us a problem. If the sections have more of a chewed up appearance, then it's probably a combination of factors: fatty tissue, not well fixed, strong antigen retrieval. We've done something for years now that seems to help. After baking, we "post fix" some of our IHC slides in formalin for about 15 minutes. Rinse in tap water then put on stainer as usual. This seems to help when certain antibodies like to chew up the tissue. We don't do this will all antibodies, mostly breast markers and those with strong antigen retrieval. I hope this helps! Good luck. Clare J. Thornton, HTL(ASCP) Assistant Histology Supervisor Dahl-Chase Diagnostic Services 417 State Street, Suite 540 Bangor, ME 04401 cthorn...@dahlchase.com From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Deloris Carter [dels...@gmail.com] Sent: Friday, July 26, 2013 5:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Losing tissue on IHC slides Hi all, I'm looking for some troubleshooting help. In the last couple of months we've been having an increasing trend in losing tissue from our IHC slides. We use a Ventana Benchmark XT. PM is done on it quarterly. The problem seems to be with the patient tissue, not the control tissue. We use Fisherbrand Superfrost Plus slides, picking up the control by dipping the top end of the slide to avoid "double dipping". We use tap water in the waterbaths which I know could cause an issue, but that's the way it's always been done here, so there's been no change to that part of the procedure. (We did try using DI water, but it didn't make any difference. Tissue still fell off of slides.) We do use DI water to mix the bulk reagents.The control slides may sometimes have been cut for a longer than recommended time, but that part of the procedure hasn't changed either, same process as always. If we run separate slides for control and patient tissue, we get much better results, but still have some tissue loss. I can't say it's all of our tests, but it is a steadily worsening problem. I'm going through tons of reagent, and I can't seem to nail down the problem. Ventana rep said to try DI water, don't double dip, try adhesive slides (which didn't really make any difference that we could tell). I would appreciate any suggestions Deloris Carter Shawnee Mission Medical Center ___ 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] Losing tissue on IHC slides
So the patient tissue and control are on the same slide, and the control is staying on but patient tissue is falling off? Maybe dont bake the control tissue just let them air dry, the humidity in the slide drying oven might be messing with the super frost plus slides. Its just a coating on the slide of some sort, i know after the first dip they dont work again like the first time, so just air dry the controls and bake after patient tissue is picked up. Sent from my iPhone On Jul 26, 2013, at 5:09 PM, Deloris Carter wrote: > Hi all, > I'm looking for some troubleshooting help. In the last couple of months > we've been having an increasing trend in losing tissue from our IHC > slides. We use a Ventana Benchmark XT. PM is done on it quarterly. The > problem seems to be with the patient tissue, not the control tissue. We > use Fisherbrand Superfrost Plus slides, picking up the control by dipping > the top end of the slide to avoid "double dipping". We use tap water in > the waterbaths which I know could cause an issue, but that's the way it's > always been done here, so there's been no change to that part of the > procedure. (We did try using DI water, but it didn't make any difference. > Tissue still fell off of slides.) We do use DI water to mix the bulk > reagents.The control slides may sometimes have been cut for a longer than > recommended time, but that part of the procedure hasn't changed either, > same process as always. If we run separate slides for control and patient > tissue, we get much better results, but still have some tissue loss. I > can't say it's all of our tests, but it is a steadily worsening problem. > I'm going through tons of reagent, and I can't seem to nail down the > problem. Ventana rep said to try DI water, don't double dip, try adhesive > slides (which didn't really make any difference that we could tell). I > would appreciate any suggestions > Deloris Carter > Shawnee Mission Medical Center > ___ > 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] Losing tissue on IHC slides
I agree water is always a potential issue. Tissue can be finicky though. Have you tried poly-L-lysine coated slides? They do help is some occasions. If you need a protocol, let me know, I can write one up, very easy. Hans B Snyder Histologistics 60 Prescott Street Worcester, MA 01605 508-308-7800 h...@histologistics.com On Fri, Jul 26, 2013 at 5:38 PM, Tony Auge wrote: > How are you drying your slides? If in an oven, what's the temp and > duration? Try going longer and tap off excess water before drying. > > > > Tony Auge HTL (ASCP) QIHC > Cell: (651) 373-4768 > Email: tony.a...@gmail.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
Re: [Histonet] Losing tissue on IHC slides
How are you drying your slides? If in an oven, what's the temp and duration? Try going longer and tap off excess water before drying. Tony Auge HTL (ASCP) QIHC Cell: (651) 373-4768 Email: tony.a...@gmail.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
