Re: [Histonet] H&E on PMMA ground sections

From:Robert Schoonhoven

I'm afraid that you are missing a critical step here.......PMMA is 
basically unaffected by any of the reagents mentioned.   A little back 
ground search through the literature, admittedly sparse as there are 
lab's making a lot of money on stents so they are not publishing 
(working out the methodology is simply time and equipment intensive, the 
science is easy), and the HistoNet archives.  I'll make my reply short 
and let you look up the specifics.  Just use a couple of 15 minute 
changes of warm xylene (55-60oC).

Robert Schoonhoven

Jason Thorsten wrote:

> Hi histonetters,
> We are embedding tissue specimens in PMMA.  We are using the EXAKT 
> system to grind the sections to approximately 25 micron sections.  We 
> are mainly working with vessels with metal implants.  We have tried to 
> stain the sections with Gills 3 Hematoxylin and Eosin Y/Phloxine.  We 
> are having problems with the nuclei taking up stain. We have tried the 
> following pretreatments before staining to no avail:
> -Treatment with 50% alcohol-2 minutes
> -Treatment with 70% alcohol-4 minutes
> -Treatement with 1%HCl in water-2 minutes
> -Treatment of 1% Ammonium Hydroxide in water-2 minutes
> -Treatment with 5% Formic acid-10 minutes
> After pretreatment, we have placed the slides in deionized water to 
> rinse quickly and placed into the hematoxylin.  We are at a loss what 
> to try.  Can anyone offer any suggestions?
> Best Regards,
> Robert Brunner
> Jason Thorsten
> RVF, Inc.
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