How do I know which version of a protocol to use for the HTL Practical?
|From:||Miriam Schroeder (by way of Histonet)|
Hello everyone. I am preparing slides for the HTL practical and am hoping
someone out there might
have some familiarity with how the slides are graded. In particular, I
need to know how to figure
out WHICH version of a stain is THE "official" version.
For example: I am requested to do an Iron stain on Bone which includes
marrow. The extent of my
instructions is this: "Iron (Prussian Blue Reaction)".
In our lab, for an iron stain, we typically follow the method listed on
page 179 of the 3rd
edition of the AFIP. This stain is called "Gomori's Method for Iron". It
uses Hydrochloric Acid,
Potassium Ferrocyanide, and NUCLEAR fast red. (And, because we have had
bad luck with Nuclear
fast red, our pathologist requests a Hematoxylin counterstain instead.)
Since that stain is not specifically called "Prussian Blue", I've been
looking further...in the
3rd edition of "Theory & Practice of Histological Techniques" (by Bancroft
& Stevens) there is a
protocol "Perls' Prussian Blue reaction for ferric iron [Perls, 1867]".
This protocol calls for
Potassium Ferrocyanide, Hydrochloric Acid, and NEUTRAL red. Can I assume
that becuase of the
specific name "Prussian Blue" that this is the proper protocol I should
follow? How can I know
(in this sort of situation in general) WHICH version of a protocol the ASCP
will want me to use
for my slides? I have a number of books I consider to be histology
"bibles" but their protocols
often vary on little details like this. Or will they not be so strict
about these aspects (like
the counterstain), as long as the Prussian Blue part is correct? How can I
be sure that a
counterstain is expected at all? Is there a "histology bible" floating
around out there somewhere
that "trumps" all others?
Any advice that can be given would be appreciated.
Also - does anyone know of a website where I could view images of what the
versions of various stains should look like?
Thanks in advance,
Research Associate, Berlex Biosciences
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