AW: [Histonet] AutoStainer XL Tweaking

[Gudrun Lang]

Hi, this is my schedule:
25 min 60°C oven - 2x5 min Xylolsubstitute - each 2 min 96%-70%-50%-A.dest.
- 10 min Mayers hemalaun - 3 min bluing in running tapwater - 45 sek. aqu.
2%eosin - - 30 sek wash in running tapwater - 30 sek 96%  - each 1 min
3x100% - 1 min butylacetat - caverslipping 

Gudrun Lang
 
Biomed. Analytikerin
Histolabor
Akh Linz
Krankenhausstr. 9
4020 Linz
+43(0)732/7806-6754
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Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Breeden,
Sara
Gesendet: Dienstag, 16. Oktober 2007 14:04
An: histonet <@t> lists.utsouthwestern.edu
Betreff: [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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