Re: Special Stains

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To:Andrea Kaj <>
Date:Wed, 12 May 1999 06:17:38 -0400
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If you go to the Histotech's Home Page

Then click on Peggy's Links to Histology Resources,

There are a bunch of links to staining procedures, histology
images, etc.

As for standardization, that's part of what the
Biological Stain Commission does. They standardize
the dry dye powders.

However, no one standardizes staining procedures
(with the possible new exception of the Her2).

I don't know if that will ever happen, as long as
people are staining by hand. First of all, many
of our stains are very "forgiving". Think of the
PAS-LG for fungus. 5 minutes in periodic acid or 10 
will give you about the same results. 10 minutes in 
Schiffs or 20? Less if heated. How is it heated?
Incubator oven? Waterbath? Microwave oven? Again, 
about the same results. 1% light green counterstain or 
0.5% or 2%? Well, we leave it on until it's the right 
shade of green. With or without hematoxylin? 
How dark? Pathologist's preference, but they could 
diagnosis either with or without, and regardless of
intensity of blue.

In addition, some of the "tweaking" that goes
on is due to differences in lab conditions.
How warm is the room? How powerful is the
microwave oven? What pH is the tap water?
Are there any impurities in the d. water?
Until these are standardized across the
country, we have to rely on the tweaking

With some of the staining machines that are
coming out (H&E, specials, IM), this may help
standardize staining. I remember when we did our
H&E by hand, and we had 8 pathologists each of
whom liked their H&E a little different. A little
more or less hematoxylin, a little more or less
eosin. So we would have to know whose slides we had, 
and stain accordingly. 

Now we have twice as many slides per day (or more), 
11 pathologists and 2 automatic H&E stainers. Now, 
all the pathologists get the same type of H&E, 
regardless of their personal likes. As long as 
the H&E is of good quality (crisp chromatin, 
several shades of eosin not overwhelming the blue), 
they all can make a diagnosis.

So, our H&E is now standardized, due to a machine.
Of course, we still had to tweak the run until it
came out to our satisfaction.

Peggy A. Wenk, HTL(ASCP)
William Beaumont Hospital
Royal Oak, MI 48073

Andrea Kaj wrote:
> Hi Netters
> Does anybody have a link to a page about special stains? I have been following the discussion about histotechnology/IHC being art or science, and it seems like we are forgetting the importance of standardising the 'old-fashioned' special stains. There is a lot of standardisation going on in IHC, but how about special stains? Any groups around?
> Some of the vendors are now offering sp. stain modules or reagents, but I am a bit concerned where they got the protocols from? And what happens when doing the stains at 60 degrees instead of room temp.? And how about disposal of various kinds of chemicals?
> Andrea
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