[Histonet] Antigen retrieval and rat skin

[Phillip Huff]

Could you give some more information about your procedure? For example, are you using frozen or paraffin sections? What thickness are your sections? One recomendation that I could provide would be to use L.A.B. Solution 
https://www.polysciences.com/shop/product.asp?pf_id=24310&dept_id=2026
which is an Ab retrieval solution we have used with success.
 
Depending on your type of section (paraffin/frozen) there are a few things I could recommend. I have also used Poly-L-Lysine coated slides with good success.
 
I would also attempt to find someone in your institution who is using the CryoJane tape transfer system. We have made some beautiful mouse skin sections as thin as 6 um that do not come off of the slide. (http://www.instrumedics.com/CryoJane.htm)
 
Phil

"Ronald P. Wilson" <rpw4 <@t> psu.edu> wrote:
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



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