[Histonet] Antigen retrieval and rat skin

[Ronald P. Wilson]

I need help! We are in the process of staining rat skin for Ki67. Ultimately
we will be counting the number of Ki67 positive cells in the epidermis. The
problem the tendency of the dermis to "blow" apart during the antigen
retrieval process. The dermal tissue either floats or folds over the
epidermis, obscuring portions of the epidermis. We have tried different
methods of heating (i.e., microwave, autoclave) and finally settled on the
Black & Decker rice steamer. The disrupted dermis seems to be less severe
with this method but still covers significant portions of the epidermis to
make counting difficult. We also tried different types of slides - plus
slides, APES-coated slides, APES-coated plus slides - and still having
problems. Any ideas?

Ronald P. Wilson
Department of Comparative Medicine, HO54
Penn State University College of Medicine
M. S. Hershey Medical Center
500 University Drive
Hershey, PA 17033
phone:  717-531-8460
fax:       717-531-5001
e-mail:  rpw4 <@t> psu.edu