[Histonet] RE: Silver stains in large batches
gayle callis
gayle.callis <@t> bresnan.net
Wed Jun 24 13:11:38 CDT 2009
You wrote:
If there is anyone out there who does silver stains in large batches I
need your input.
I haven't done a Sevier-Munger stain in probably 20 years and when I
did this kind of stain I usually had 2-3 slides to stain. So now I
have a request to do this stain and I have multiple racks of slides.
What is the best way to handle the ammoniacal silver step - each slide
individually or plunk a whole rack of slides into a staining dish? And
if I do the latter, can I multiply the amounts by 5 to make 250 ml. of
the ammoniacal silver solution which is the amount needed to fill the
staining dish?
Thanks
Andi
**********************
Andi,
I did Steiner and Steiner Helicobacter staining in huge batches, 20 slides
per rack to control the timing better. I used glass staining dishes (metal
is unacceptable for silver stains) and crisscrossed the slides so they were
not back to back. Criss crossing slides avoids trapped reagents that cause
nasty carryover. I had a positive control in EACH rack to monitor the
silver deposition for that rack of slides. I monitored with a microscope
and you may need to do this too. It took a bit of careful planning and with
heated solutions for Steiner another dimension of difficulty. I locked
doors and didn't answer telephones. The staining worked beautifully, and I
could do 60 or more slides a day. Mass production is possible and if it was
me doing this, there is no way I would do 2 to 3 slides at a time. It would
be a nightmare workday, maybe an "all nighter" in the lab!
Good luck
Gayle M. Callis
HTL(ASCP)HT,MT
Bozeman MT 59715
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