Re: [Histonet] alcian blue pH

From:Geoff McAuliffe

Hi Tom:

    Alcian Blue will stain the glycosaminoglycans in the extracellular 
matrix, it won't show much cellular morphology. Still, the traditional 
method is 1% Alcian Blue in 3% acetic acid for a pH of 2.5. At 2.5 both 
sulfated and carboxylated GAGs are stained. A red nuclear stain, Scarba 
red or nuclear fast red, gives a nice contrasty counterstain.
    You might try 0.05% Toluidine blue in dist water with a pH of about 
4.0 for 10-15 minutes, dist water rinse and dehydration in 3 changes of 
actone followed by clearing in fresh xylene. Nice metachromasia of=20
extracellular matrix and you will see some cellular morphology as well.


Tom Schaer wrote:

>Greetings List:
>I would like to stain some intervertebral disc x-sections (rat, decalcified) with alcian blue.  Interest in cellular morphology of nucleus pulposus, transition and annulus cells.
>What pH of the stain should I use.
>Many thanks, 
>Thomas P. Schär, VMD
>Asst  Prof  School of Biomedical Engineering, Drexel University
>& Comparative Orthopaedic Research & Tissue Engineering
>Department of Clinical Studies
>University of Pennsylvania New Bolton Center
>382 West Street Road
>Kennett Square, PA 19348
>tel. 610-444-5800 (x6261 office)
>tel. 610-444-5800 (x6131 lab)
>fax. 610-925-8100
>Histonet mailing list

Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029

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