RE: Cartilage, Alginate, and Differential Staining
Takeuchi's acriflavine (Stain Technology 37:105-109; 1962) usually
distinguishes between sulfated polysaccharides and uronic acids.
Spicer & Meyer's aldehyde fuchsin-alcian blue sequence (Am.J.Clin.Pathol.
33:453-460; 1960) almost always distinguishes between sulfated
polysaccharides and uronic acids.
Allen A. Smith, Ph.D.
School of Graduate Medical Sciences
Podiatric Medicine and Surgery
Miami Shores, Florida 33161-6695
From: Tracey Couse [mailto:firstname.lastname@example.org]
Sent: Friday, April 19, 2002 10:05 AM
To: HistoNet Server
Subject: Cartilage, Alginate, and Differential Staining
I was hoping someone could provide some insight to my present dilemma. Let
me provide some background. I have been given some alginate gels seeded
with chondrocytes. They were formalin fixed and paraffin embedded. I have
been asked to stain for cartilage. Initially this sounds easy, but the
problem is that the alginate gel tends to stain with similar properties to
that of the cartilage of interest. It becomes very difficult to
differentiate between the gel and the cartilage. There are some differences
in staining intensity, but the gel is not uniform in density so it too
stains with different intensities masking the cartilage.
I have learned that the alginate gels are comprised of linear polymers of
guluronate and mannuronate. I would have thought that because the
glycosaminoglycans (gags) contain sulfates, that I would have been able to
use alcian blue to differentiate between the sulfates of the gags and
carboxylates of the alginate. This has not been the case. To date, I have
tried staining with safranin-o, toluidine blue, and alcian blue at pH 1 and
pH 0.5. Again, the results have been that the alginate gel takes on roughly
the same shade of stain as the cartilage (I am using articular cartilage
for a positive control).
Any suggestions, tips, ideas, corrections, and/or knowledge would be
greatly appreciated. I am new to the cartilage and bone field so any
insight from the HistoNet would be invaluable to me.
Georgia Tech/Emory Center for the
Engineering of Living Tissues
Georgia Institute of Technology
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