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Journal of Histochemistry and Cytochemistry, Vol. 46, 1455-1460, December 1998, Copyright © 1998, The Histochemical Society, Inc.

TECHNICAL NOTE

Detection of Antigens by Immunofluorescence on Ultrathin Cryosections of Skin

Akira Ishikoa,b, Hiroshi Shimizua, Takuji Masunagaa, Yukiko Kuriharaa, and Takeji Nishikawaa
a Department of Dermatology, Keio University School of Medicine, Tokyo
b Department of Dermatology, Nippon Kokan Hospital, Kawasaki, Kanagawa

Correspondence to: Akira Ishiko, Dept. of Dermatology, Keio Univ. School of Medicine, 35 Shinanomachi, Shinjuku, Tokyo 160-8582, Japan..

Cryoultramicrotomy was originally established to provide ultrathin cryosections as substrates for on-section immunolabeling in immunoelectron microscopy. Recently, we recognized that ultrathin cryosections of skin (0.2 µm thick) could serve as substrates for immunofluorescence (IF) with excellent resolution. To assess the advantages and the limitations of IF on ultrathin cryosections, we compared the labeling of IF on 0.2-µm ultrathin cryosections of skin with those of routine IF on 6-µm cryostat sections, confocal laser scanning microscopy (LSM), and immunogold electron microscopy using several markers of keratinocyte cell surface and basement membrane zone molecules. IF on ultrathin cryosections clearly demonstrated a lack of bullous pemphigoid antigens beneath the melanocytes, desmosomal antigens as discontinuous dot-like labeling, and nondesmosomal plasma membrane antigen as a ladder-like pattern. IF on ultrathin cryosections provided convincing images with higher resolution than confocal LSM, which corresponded well to those of immunogold electron microscopy. IF on ultrathin cryosections had superior resolution compared to routine IF or confocal LSM and should serve as a powerful tool in future studies for the analysis of skin antigens. (J Histochem Cytochem 46:1455–1460, 1998)

Key Words: immunohistochemistry, basement membrane zone, desmosome, hemidesmosome, confocal laser scanning microscopy, cryoultramicrotomy


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