[Histonet] PAS staining

[LuAnn Anderson]

  I still make my own Schiff's reagent. It is easy to make and I've 
never had a problem with it. Keeps well in refrigerator. I do make the 
periodic acid fresh each time. If anyone wants the recipe, let me know.

LuAnn


On 10/6/2010 11:43 AM, Janet Keeping wrote:
> I have for some time had a problem with Schiff's reagent and PAS staining.
>
>     - I have tested each new, unopened bottle of Schiff's reagent with
>     formaldehyde and always the color development was immediate, but purple,
>     definately not pink.This result has been quite consistant.
>     - PAS staining for glycogen using the method in *Histotechnology a self
>     Instructional text,* would fail to demonstrate any glycogen in autopsy
>     liver specimens.
>
> I teach Histology at a community college and this problem has driven me
> crazy for a number of years. I have tried several brands of Schiff with the
> same results. Recently I obtained sheep tissues which were promptly
> refrigerated and fixed after death, and I had hoped these tissues would
> demonstrate glycogen. ( My thinking was that perhaps delay before autopsy
> was somehow diminishing the glycogen in the specimens that I had.or that
> perhaps long term NBF fixation had hampered staining.) Basement membranes
> were stained with the Schiff reagent as expected despite the purple color in
> the formaldehyde test. Hotchkiss Mcmanus with the same reagent also produced
> beautiful staining of fungus a lovely magenta color. A search of the web
> made me suspiciious when I noted that Schiff added to 37-40% formaldehyde
> should produce a pink or red color, however, A spot check of formalin using
> Schiff should produce a purple color. I sent an e-mail to Brian Hewlett to
> ask if he could make any recommendation.
>
> Brian was not surprised by the purple color devopment in testing, He
> suggested that a large number of available aldehyde groups would be expected
> to produce this color. He suggested that I increase my periodate oxidation
> to 20-30 minutes and my Schiff application to 30 minutes.
>
> This worked and I am extremely grateful!. Has anyone else had an experience
> like this?
> Janet
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