[Histonet] Tissue Washing

[Kemlo Rogerson]

Have you changed your Supplier for glass slides, has your Supplier
changed theirs? Have you poorly quality slides, are they dirty, are they
greasy. Have you tried cleaning them in alchohol by soaking and then
rubbing with a cloth? If all else fails albuminise the slides but you
might find that the albumin stains up too.

>From my experience it usually the slides and as it has been happening
for some time you may like to change to a better quality glass slide;
make sure that they are not 'old' slides, you know old stock? Water OK
in the water baths? 


 
Kemlo Rogerson	
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Emily
Butte
Sent: 31 July 2009 14:58
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Tissue Washing

Our lab has been struggling for a while now, with tissue washing off the
slides. The problem was originally traced to our processor that was
malfunctioning and having major carry-over issues, causing poor tissue
processing. So after failed attempts to get the old machine fixed, we
ended up getting a new microwave processor. The tissue (mainly prostate
bx) comes out of the processor looking good. It cuts well, but after
staining, some of the sections are no longer on the slides. The problem
was quiet severe with the old processor and has decreased drastically,
but has not entirely gone away with the microwave processor.

Here's some info on our current procedure. This is a small physician
office lab and we everything manual. We use plus slides and a water bath
with no tissue adhesive added. The bath is between 42-45 degrees
Celsius. After sectioning, the slide are put in an oven at 60 degrees
for 20 minutes. After taking the slides out of the oven, we cool them to
room temp then hand stain them in an H&E line.
Xylene - 5 min
Xylene - 2 min
Xylene - 2 min
100% Alcohol - 30 sec
100% Alcohol - 30 sec
100% Alcohol - 30 sec
95% Alcohol - 30 sec
Rinse in DiH2O
Hematoxylin - 2 min
Rinse in DiH2O
0.50% Acid Alcohol - 2 dips
Rinse in DiH2O
Scott's Tap Water - 10 sec
Rinse in DiH2O
95% Alcohol - 30 sec
Eosin - 30 sec
95% Alcohol - 30 sec
100% Alcohol - 30 secs
100% Alcohol - 30 secs
100% Alcohol - 30 secs
Xylene - 30 sec
Xylene 2 min
Xylene - 2 min

The slides are then cover slipped and of a 12 piece prostate case, at
least a section or 2 will wash off of about 4 of the slides. Please pass
along any ideas as to what the cause of this could be and how I can fix
it. I'm fresh out of ideas and would really appreciate the advice of
someone with more experience than myself.

Thanks,
--
Emily
Path Lab Solutions Inc.
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