Re: [Histonet] HYALURONIC ACID detection
Some properties of hyaluronic acid.
(1) Stains with alcian blue 8G at pH 2.5
but not at pH 1.0.
(2) Staining prevented by prior treatment with
streptococcal hyaluronidase. (Testicular
hyaluronidase is cheaper & more easily
available but can also remove chondroitin
sulphates. Chondroitin sulphates are
stained by alcian blue at both pH 1 and 2.5.
(3) Staining by alcian blue is reversibly
blocked by prior methylation of the section.
Stainability is restored if methylation is
followed by saponification. The staining of
chondroitin sulphates with alcian blue is
irreversibly blocked by methylation.
(4) Negative with classical PAS method but
becomes PAS-positive if a special
oxidation sequence is used (Scott &
Harbinson 1969 Histochemie 19:155-161).
If you have pure hyaluronic acid there will
be no difficulty staining it. If, as is usually
the case, other macromolecular carbohydrates
or glycoproteins (collagen, for example) are
present in exactly the same place, you'll need
to do all the above methods and more. The best
account of carbohydrate histochemistry that I
know is Chapter 15 in Vol 2 of Pearse's
"Histochemistry". It's quite heavy going,
but this is a field where you really do need
to be in command of every step of every method.
Hope this helps.
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London, Canada N6A 5C1
Hernan Aldana wrote:
> Dears histonetters
> I like to demonstrate the presence of hyaluronic acid in fixed eyes. Can
> anybody help me with some advices?
> Histochemical or immunohistochemical. Please help me.
> Thanks in advance to all
> Dr. Hernán J. Aldana Marcos
> Facultad de Medicina. Universidad de Morón
> Machado 914. B1708JPD. Buenos Aires. Argentina
> e-mail alternativo firstname.lastname@example.org
> web: http://hjaldanamarcos.bravepages.com
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