Hi there,

Perhaps it's the depth when originally mounting the tissue on the
slide.  Do you use some sort of Sta-On in your waterbath?  It could be
the cause of retaining excess hematoxylin.  Barb

>>> "Molinari, Betsy"  08/14/06 12:22PM
>>>
Bonnie,
It is horizontal. I thought the same thing..reagent depth..so I sat
and
watched the entire run to be sure the basket/slides were submerged and
they were. I truly appreciated this help, it is frustrating to be
looking at another tray of slides w/ streaks.

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: Bonnie Whitaker [mailto:bwhitaker@brownpathology.com] 
Sent: Monday, August 14, 2006 11:17 AM
To: Molinari, Betsy
Subject: RE: [Histonet] Leica DRS

Betsy,

Where is the blue streak?  Is it vertical or horizontal?  Is it always
the
same place?  I'm wondering if it's a depth of reagent issue (tissue
isn't
totally submersed in all steps).

Bonnie Whitaker


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu 
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
Molinari,
Betsy
Sent: Monday, August 14, 2006 10:54 AM
To: histonet@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)

 

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