Re: [Histonet] Movat's Pentachrome question

From:"Anthony Reilly"

The alcoholoic ammonia step is the critical stage of Movat's as it is 
required to convert the Alcian Blue to insoluble Monastral Blue 
so that it will not be removed during the remainder of the
 staining method.  Just follow the AFIP method which uses 10ml of
ammonium hydroxide in 90ml of 95% alcohol.
If you want to simplify things then you can do what we do.  Just 
stain using your usual Alcian blue method(pH2.5), treat with alcoholoic
ammonia for 2 hours then do a routine VVG.  This will stain your elastic
fibres, collagen and glycoproteins as the Movat does with  routine 
stains that are already on your shelf for other methods.

Good luck

Tony Reilly
Chief Scientist
Anatomical Pathology
QHPS-Prince Charles Hospital
Rode rd Chermside Q 4032
Ph: 07 3350 8543
Fax: 07 33508546

>>> "Monfils, Paul"  11/30/05 4:52 am >>>
Does anyone do Movat's Pentachrome stain?  I have a researcher who wants to
do it.  My question involves the "alkaline alcohol" the procedure calls for.
The protocol says to add ammonium hydroxide drop by drop to 95% ethanol
until the pH reaches 8.0 or higher.

Question 1, a theoretical one - How can you accurately measure pH in a
nearly water-free solution?  Presumably an ionic compound would not ionize
very well in such a solution?

Question 2, a practical one - When I put the pH probe into 95% ethanol, the
meter already reads higher than 9.0, even without adding any ammonium
hydroxide. Is this your experience?

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