Hi Sara,
Here is my schedule for H&E. It's not the Leica stainer but Microm
DS50.
Maybe this will help.
Baseball in Denver at the NSH??
Linda
1 Oven 15 minutes
2 Xylene 3 minutes
3 Xylene 3 minutes
4 Xylene 3 minutes
5 100 % Alcohol 1 minute
6 100 % Alcohol 1 minute
7 100 % Alcohol 1 minute
8 95% Alcohol 1 minute
9 95% Alcohol 1 minute
10 Running Water 2 minutes
11 Hematoxylin 3 minutes
12 Running Water 2 minutes
13 5% acetic acid 15 seconds
14 Running Water 1 minute
15 Bluing Reagent 1 minute
16 Running Water 1 minute
17 95% Alcohol 15 seconds
18 Eosin 2 minutes
19 100% Alcohol 15 seconds
20 100 % Alcohol 1 minute
21 100 % Alcohol 1 minute
22 100 % Alcohol 1 minute
23 Xylene 2 minute
24 Xylene 2 minute
25 Xylene 2 minute
Linda Blazek HT (ASCP)
Manager/Supervisor
GI Pathology of Dayton
7415 Brandt Pike
Huber Heights, OH 45424
Phone: (937) 293-4424 ext 7118
Email: lblazek@digestivespecialists.com
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Breeden,
Sara
Sent: Tuesday, October 16, 2007 8:04 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] AutoStainer XL Tweaking
I'm setting up my Leica AutoStainer XL and would appreciate any help
with tweaking the times. I based my timing for the Autostainer on my
manual stain times but staining is not crisp and looks washed out. Docs
say eosin is fine but cellular detail is foggy. Here are my times:
XYL - 3 at 5 min each
100% reagent - 2 at 30 seconds each
95% reagent - 1 at 30 seconds
80% reagent - 1 at 30 seconds
DI water wash - 1 minute
Hematox (currently using Sigma) - 5 minutes
DI water wash - 1 minute
Acid Alcohol (1%) - one dip
DI water wash - 1 minute
Ammonia water - 15 seconds
DI water wash - 1 minute
80% reagent - 1 at 30 seconds
95% reagent - 1 at 30 seconds
Eosin (alcoholic) - 10 seconds
95% reagent - 1 at 30 seconds
100% reagent - 2 at 1 minute each
XYL - 3 at 1 minute each
I cut my acid alcohol time down from 15 seconds to one dip but it's
still taking a lot of hematoxylin off. Should I try 0.5% acid alcohol?
I'm at a loss so far; different hematox? More time in ammonia water? If
you use this Leica AutoStainer XL, could you offer some Handy Hints? I
thank you SO much!
Sally Breeden, HT(ASCP)
NM Dept. of Agriculture
Veterinary Diagnostic Services
PO Box 4700
Albuquerque, NM 87106
505-841-2576
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