[Histonet] PGP 9.5 for intraepidermal nerve fibers

[MaryAnn Dixon]

Hi Netters,

 

I'm stumped with a project and could really use your help. I have 3mm
punches of skin that I need to stain with PGP 9.5 but I am specifically
looking for the intraepidermal nerve fibers.  The sections were given to
me fixed in formalin and then I made cryo buttons out of them using OCT.
I have found various articles that have cut sections ranging from 4
microns on up to 50 microns. I cut a slide to try at 20 microns but need
help after this.  I am using a polymer based detection system and the
antibody is polyclonal from Dako.  I know the antibody works well on
paraffin embedded tissue but I'm lost with a starting point for these
frozen 20um sections.  Some journal articles also have the primary
incubating overnight. Is this the only way I will be able to see the
IENF's? If anyone out there is doing this kind of work can you please
contact me. Thank you in advance.

 

 

MaryAnn Dixon  BS

Biological Scientist

Anatomic Pathology

UF Veterinary Medical Center

(352) 392-2235 Ext. 4517