[Histonet] Stripped H&E's - HELP!

[Sheila Fonner]

Jo-Ann,

I have seen this problem many times over the years.  Nearly 100% of the time
it is due to the level of the alcohol after the eosin being too low.
Whether you water rinse or go straight into a 95% etoh, the level of your
first alcohol after eosin MUST be covering your entire slide.  If not, the
eosin will slowly drip down the slides in the subsequent solutions.  This
can also happen after hematoxylin.  Keep the hematoxylin and eosin LOWER
than the solutions that come after them.  Hope this helps.

Regards,
Sheila
KDL Pathology
Knoxville, TN


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jo-Ann
Bader, Ms.
Sent: Thursday, July 07, 2011 3:38 PM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Stripped H&E's - HELP!

Hi All,

Over the past year we have had a recurring problems with our H&E's.  The
come out of our Gemini Stainer stripped but not on every run; we can go for
months without a problem and then all of a sudden it is back again.  The
stripping is in the Eosin, our pathologist confirms the the hemotoxylin is
not affected.  There are 3 of us cutting on 3 different microtomes.  The age
on the microtomes range in age from 10 years to less than 1 year. 

Some of the solution we have tried are:

We numbered all our racks to see if some of our older racks may have micro
cracks in them that may retain solution which gradually dripped onto the
slides.  After checking for over a month there was no pattern to  racks and
bad staining.

We tried lowering the level of the solutions to below the coated end of the
slides.  No results

We tried changing the positions of the pots for the solutions,  thinking
that if there was a leak from the solutions above we would notice a
difference.  No luck.

We checked to see if the stripes only appeared near the end of our stains
life.  Not that either

Right now I have a technician sitting in front of the stainer as it runs to
see if she can catch any abnormalities in the way the machine is
transferring the slides.

We rotate our alcohols every run.

We change every solution after every 450 slides.

I have put a call into the company to see if I can get to speak to a
specialist on this machine but in the mean time I was hoping someone out
there could help.  Has anyone seen such a problem

I, we, have run out of ideas aside from it being the machine and not being
able to find out exactly how.


Jo-Ann Bader
Histology Co-Ordinator
Goodman Cancer Centre
McGill University 1160 Pine Ave. W - Rm 312 (3355)
Montreal, QC, Canada
H3G 1Y6
Tel: 514-398-8270
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet