[Histonet] Leica DRS

[Molinari, Betsy]

You know what..DUH! I the stripe is vertical. I did not consider how the
slide is positioned while in the stainer which is vertical. So the drop
running down the slide sounds sensible. How limited was my perception of
this problem? OK..fresh eyes..new ideas..thanks!

Betsy Molinari HT (ASCP)
Texas Heart Institute
Cardiovascular Pathology
6770 Bertner Ave.
Houston,TX 77030
832-355-6524
832-355-6812 (fax)


-----Original Message-----
From: Morken, Tim - Labvision [mailto:tpmorken <@t> labvision.com] 
Sent: Monday, August 14, 2006 11:31 AM
To: Molinari, Betsy
Subject: RE: [Histonet] Leica DRS

Betsy, you don't say which direction the streak goes, but I am guessing
it
is vertical (on the long axis of the slide). If this is the case I
suggest
you look at the level of your eosin compared to the 95% just before it.
We
had this problem once and discovered the eosin level was lower than the
95%
level in the previous bath. Then when the slide went into eosin it did
not
rinse off the 95% at the top and left a drop above the level of the
eosin.
Then when the slide came out of the eosin that drop would slide down the
middle of the slide and wash out some of the eosin before going into the
next alcohol. It doesn't sound like much, but was enough to make a
"blue"
streak down the slide. It was really some of the eosin washing out.

Tim Morken
Lab Vision - Neomarkers
www.labvision.com



-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Molinari,
Betsy
Sent: Monday, August 14, 2006 8:54 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Leica DRS


Hi,

I am frustrated. I have been struggling to solve this problem and now
turn
to you.

My H&E's  come out of the stainer with this blue line across them. I
have
juggled times, rinses, solutions.

The slides are cut at 4-5 microns. This is my current protocol

Dryer: 20 min

Xylene x3 5 min

100 % x3 1 min

95% 1 min

rinse in tap

rinse in DH2O

 Richard Allan hematoxylin 1  1 min ( initially started w/ hematoxylin 2
but
staining was too dark, I have juggled these times from 15 secs-1
min)

Rinse in tap 2 min

Richard Allan Clarifier 2  30 secs (have played w/ this time too)

Rinse in tap 2 min

Richard Allan Bluing 1 min

Rinse in tap  2 min

95 % 1 min 

Richard Allan Eosin Y 45 secs

100%- Xylenes  I min.

 The staining is acceptable..good nuclear staining but still that damn
blue
streak! Any suggestions?

Thanks.

 

Betsy Molinari HT (ASCP)

Texas Heart Institute

Cardiovascular Pathology

6770 Bertner Ave.

Houston,TX 77030

832-355-6524

832-355-6812 (fax)

 

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet