[Histonet] Autostaining issues

Breeden, Sara sbreeden <@t> nmda.nmsu.edu
Wed Jan 2 15:46:53 CST 2008

Let me be as succinct as possible without installing a prejudice to the
answer. I have an automatic stainer. I had a DI system installed with
the explicit purpose of preventing calcium build-up in the stainer and
causing me pain and agony in keeping it de-crusted. I took the manual
stain line solutions/times and translated it to the stainer; I have
tweaked and fiddled but the stains look as if they were done on necrotic
or autolytic tissues! So I pondered reasons and fiddled/tweaked more. I
hit on pH- I checked the DI water (pH 5.12!!), and the tap water (pH
7.98). I thought that was the solution to my problem, so I cut back on
time in acid alcohol and cut back in wash times.  No luck. Changed
hematoxylin vendors. Nope. Made up 0.5% acid alcohol. Nada. Increased
time in ammonia water. Not that either. The stainer vendor has been very
accessible but we have not yet solved the poor staining - and it's NOT
the stainer's fault. I've been on the phone with the DI cartridge
supplier and he's telling me that pH may not be a factor because the DI
filter adds elements (sodium?) but removes elements (calcium) and that
the water contaminates the reading anyway. So I'm turning to my Panel of
Experts. Is it pH, as I suspect? Is it something else? I'm facing a
large workload for a project this spring and would like to use the
autostainer - as well as be able to use it for the daily surgicals and
necropsies. I know there's someone out there that can help me figure
this out!  Thanks!


Sally Breeden, HT(ASCP)

NM Dept. of Agriculture

Veterinary Diagnostic Services

PO Box 4700

Albuquerque, NM  87106



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