Re: [Histonet] Alcian Blue deposit
If an alcian blue solution contains insoluble
material, or if insoluble material forms after
filtering, the solution must be no good. Solutions
at pH 1.0 or pH 2.5 can be stable for some years.
Solutions of alcian blue used in "critical
electrolyte concentration" procedures are stable
for only a few hours. See Horobin 2002 - Ch.26 in
Conn's Biological Stains, 10th edn.
Insoluble blue material is almost certainly copper
phthalocyanine pigment. The books say this
dissolves in concentrated sulphuric acid, but it
can't be removed with any ordinary solvent.
Alcian blue has been available as a certified dye
powder since 1981, so use only a certified batch.
The solid dye powder can sometimes deteriorate;
this may be related to dye purity, with certain
non-dye additives (notably boric acid) improving
stability. Generally, solid dyes are stable for
A related dye is the alcian blue pyridine variant
(Sigma-Aldrich), which is more stable and reliable
than alcian blue (see Churukian, Frank & Horobin,
2000 Biotech. Histochem. 75: 147-150; Henwood 2002
Biotech. Histochem. 77: 93-94.) With the pyridine
variant you do not obtain a completely
acid-resistant product like that produced by
staining with real alcian blue, but that may not
matter for many purposes. Another more stable
alcian blue substitute is alcec blue; I don't know
if this is still available. The most recent alcec
blue publication (of very few) that I've seen is a
nicely illustrated 2001 paper on sulfoglycolipid
storage disease by Lullmann-Rauch et al, in
Histochemistry & Cell Biology 116:161-169.
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London, Canada N6A 5C1
SARAH REEVES wrote:
> Does anyone have any ideas to reduce alcian blue deposit on sections. We filter the solution after making up as well as immediatly before use.
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