[Histonet] double staining for nerve fibers and endothelial cells on skin samples

From:"Cao, Xudong"

Dear all:
I am trying to determine the time and spacial sequence of nerve innervation and angiogenesis of a grafted skin equivalent using histo-chemical-immunostaining. The primary antibodies that I will be using for this study are PGP9.5 for nerve fiber and CD-31 (i.e. PECAM-1) for vascularization, and I will be using two different but compatible peroxidase substrate to visualize the signals . My questions: 1) is there anything that I need to pay special attention to for double staining in general? 2) the staining for nerve fiber normally calls for a thick section (40 to 50 um) whereas that for endothelial cells calls for a relatively thin section (6-8 um). How can I accommodate this difference in thickness, any suggestions? thanks for the help.
Xudong Cao, Ph.D.
Post-doctoral fellow
Brown University

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