That could be the solution. The problem is that you changed 2 things: the protocol and the slides and now you do not know which is "the culprit". Returning to the old protocols is a first step to trying to solve the present situation and, as you can imagine, if the old protocol does not solve the problem, the slides may be the cause. René J.
--- On Thu, 2/2/12, Sherwood, Margaret <msherw...@partners.org> wrote: From: Sherwood, Margaret <msherw...@partners.org> Subject: RE: [Histonet] slides for frozens To: "Rene J Buesa" <rjbu...@yahoo.com>, "histonet" <histonet@lists.utsouthwestern.edu> Date: Thursday, February 2, 2012, 3:03 PM That may be true, but I don't recall having this problem with our previous protocol (which was longer and included a fix in alcoholic formalin). I think we will try the "old" method and see what happens. Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (EDR 214) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114-2696 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-1206 (fax) msherw...@partners.org ________________________________ From: Rene J Buesa [mailto:rjbu...@yahoo.com] Sent: Thursday, February 02, 2012 2:57 PM To: histonet; Sherwood, Margaret Subject: Re: [Histonet] slides for frozens There is a known saying that goes: "You get what you pay for". René J. --- On Thu, 2/2/12, Sherwood, Margaret <msherw...@partners.org> wrote: From: Sherwood, Margaret <msherw...@partners.org> Subject: Re: [Histonet] slides for frozens To: "histonet" <histonet@lists.utsouthwestern.edu> Date: Thursday, February 2, 2012, 2:46 PM To all: We have been using the same slides for paraffins and frozens. We have no trouble with paraffin-embedded tissue adhering to the slides through the whole staining procedure. But our frozens (even 5um sections)have been falling off during manual staining. The slides are (+) charged. We used to use slides from Fisher, but they are so expensive, we changed to an independent vendor (half the price). Our procedure for frozen H&Es has recently changed: 1. Fix in 95% ETOH 1 min. 2. Hematoxylin 5 min. 3. Acetic acid H2O 2 quick dips 4. Wash-running H2O 10 seconds (10 dips) 5. Bluing reagent 10 dips 6. Wash-running H2O 10 seconds (or 10 dips) 7. Eosin 5 seconds 8. Dehydrate-95% ETOH 10 dips each 9. Dehydrate-100%ETOH 10 dips each 10.CitriSolv 10 dips each 11.Coverslip The tissue seems to fall off in the bluing reagent (we order from Mossberg). Can anyone help us? Suggest anything--what slides you are using? We have had the same problem with special stains (i.e. Oil Red O for fat on frozens). Thanks! Peggy P.S. I use the same slides for my 1um plastic embedding which I dip in water rinses and have no problem with the tissue falling off. Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (EDR 214) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114-2696 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-1206 (fax) msherw...@partners.org <http://us.mc1621.mail.yahoo.com/mc/compose?to=msherw...@partners.org> 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://us.mc1621.mail.yahoo.com/mc/compose?to=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