RE: Reticulin stain
|From:||"Monson, Frederick C." |
The first time I made up AFS (ammonium ferric
sulfate)[(a la, Hdb Chem & Physics, 63rd))] I was told to carefully pick
only the lavender crystals (violet in the above book for the compound identified
as: "ammonium iron sulfate(ic)" [NH4Fe(SO4)2.12H2O] and not those that
were white. The latter are identified as "ammonium iron(III) sulfate"
[(NH4)2SO4.Fe2(SO4)3]. How the one converts to the other with age in the
jar was not explicitly stated and remains unknown to me, but the admonition was
VERY clear, if I wanted the reticular or iron hematein stains to work.
While students have departed and lost, it is one area in which I and my mentor
have always been in absolute unbending agreement. [For the reason, that I
have never seen any evidence to refute the original
use only the lavender (violet) crystals, the solution will have a color that
will approximate dilute urine, that is, a
light, delicate yellow hue.
pure aside, I once retrieved a container in which all the crystals appeared to
be white, and even though the white is less soluble in cold water than the
lavender(violet) [44:124/L]. I retrieved about 250ml worth of the larger
crystals, placed them in a double thickness of #2 Whatman filter paper, and ran
a small amount of vigorously boiling water over them to see if I could remove
the white and be left with any lavender. I recovered just enough before
they all dissolved.
Finally, yes, I have filtered solutions of AFS(FAS) or whatever, and felt
better. On the other hand, if I make the solution in the morning for an
afternoon procedure, the little flecks of stuff are no longer floating and thus
not bothersome when I decant from above them.
when I first tried the Gordon and Sweet as represented by Pearse in 1960(?), I
was prewarned, by the author, about the vagaries of the procedure and the
variety of 'color' in the result. Undaunted, I went forward, if only to
finally use a method that beautifully counteracted the constant odor of HCHO in
the anatomy lab in which I worked. Gordon and Sweet occupy a lone position
at the top of my "most awakening/stimulating" histologic methods. All the
others remain interesting, but only for the results that they
ordered "Ammonium iron sulfate(ic)"(?), I always demanded that it be from a
fresh lot and have, on opening, nothing but lavender crystals. If the lots
were fresh, the crystals were as they should have been.
is correct about the other types of collagen, and you will not be able to speak
of "Type III" without doing an adjacent, frozen section with an appropriate
Ab. With respect to Ab's, however, it is still not absolutely clear that
an Ab against Type III or Procollagen will 'show' all so-called reticular
fibers. It appears to be an interesting non-conjunction of
histology and immunohistochemistry - unless, of course, someone
will educate me different!
are welcome, and come back anytime.
Hope this helps and gives cheer,
Frederick C. Monson, PhD
Center for Advanced Scientific
Mail to Geology
West Chester University of
Schmucker II Science Center, Room SS024
South Church Street
and Rosedale Avenue
West Chester, PA, 19383
am preparing solutions for a reticulin stain. This is my first time
using Ferric Ammonium Sulfate, or doing a reticulin stain. When mixing
Ferric Ammonium Sulfate in Distilled Water is the solution supposed to be
Rust/Light orange colored with a white precipitate that will not
dissolve? If so is this solution supposed to be filtered? Any
other recomendations would be appreciated, thanks for your time.
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