Ahhh!!! I had this same problem when I came to this lab too!! Plus if you put that xylene substitute crap (mine was clear rite) it screws up cell blocks that have been embedded into agar!! 5 experiments had to be repeated because of this stuff!! The eosin bleed is because you got water on the slides after the eosin. With xylene you can see the water pool at the bottom, but with these substitutes it just kind of becomes part of the solution. I have finally gone back to xylene! We had to buy hoods, but it's the cost of better histology =)
Fixing the eosin bleed...uncoverslip, run through xylene (or let sit in the substitute for 15 or 20 minutes, then alcohols, then in running water for 10 minutes or so. Then change all your alcohols to fresh solutions (I would change the substitutes too), then from water to alcohol, eosin, etc. etc. and they should be fine. Good luck!! I wanted to pull my hair out a couple months ago because of that stuff...run away from it!!! Sarah Goebel-Dysart, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Christopher Conlisk Sent: Friday, June 03, 2011 1:18 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Slide Bright Question Hello Everyone, I have worked in labs that use Xylene my entire career and I just started at a Lab That only uses a Xylene Substitute "Slide-Bright". I am having problems with the H&E. After staining and coverslipping (The slides look fine innitially), Then about 5-10 minutes after coverslipping the Eosin starts bleeding out all around the tissue. I have asked several of my Histotech Friends that are old timers and they say that Xylene Substitutes are awful at deparrifinization and awful at clearing. They told me that the alcohol isnt getting thoroughly cleared in the "Slide Brite" and then it is eventually leeching out after coverslipping??? Is this true and does anyone have any guidance for this issue? We also run MOHS slides on the same stainer and I keep all the reagents clean as a whistle. I really hate Xylene Substitute's.... Thanks C.S. Conlisk HT(ASCP), PBT(ASCP) Kansas City Skin and Cancer 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