[Histonet] AutoStainer XL Tweaking

[Blazek, Linda]

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 <@t> digestivespecialists.com

 


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Breeden,
Sara
Sent: Tuesday, October 16, 2007 8:04 AM
To: histonet <@t> 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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