[Histonet] Slide Bright Question

[sgoebel <@t> mirnarx.com]

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-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Christopher Conlisk
Sent: Friday, June 03, 2011 1:18 PM
To: histonet <@t> 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 <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet