[Histonet] Movat's Pentachrome question

[Lori Richey]

We do the Movat's stain.  The Alkaline alcohol recipe that we use is 45 
ml 95% ETOH, 5 ml Concentrated Ammonium Hydroxide.  We preheat, the 
alkaline alcohol sol'n in a 60 degree oven ( I know this is very 
flammable), and stain in preheated sol'n for 10 min.  We don't pH at all.

Anthony Reilly wrote:

>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
>regards
>
>Tony Reilly
>Chief Scientist
>Anatomical Pathology
>QHPS-Prince Charles Hospital
>Rode rd Chermside Q 4032
>Australia
>Ph: 07 3350 8543
>Fax: 07 33508546
>tony_reilly <@t> health.qld.gov.au
>
>
>  
>
>>>>"Monfils, Paul" <PMonfils <@t> Lifespan.org> 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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