[Histonet] H&E on PMMA ground sections

[Jason Thorsten]

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
rbrunner <@t> rvfarms.com

Jason Thorsten
jthorsten <@t> rvfarms.com

RVF, Inc.