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Journal of Histochemistry and Cytochemistry, Vol. 48, 481-486, April 2000, Copyright © 2000, The Histochemical Society, Inc.

SYMPOSIUM

FluoroNanogold Is a Bifunctional Immunoprobe for Correlative Fluorescence and Electron Microscopy

Toshihiro Takizawaa and John M. Robinsonb
a Department of Anatomy, Jichi Medical School, Tochigi, Japan
b Department of Physiology and Cell Biology, Ohio State University, Columbus, Ohio

Correspondence to: Toshihiro Takizawa, Dept. of Anatomy, Jichi Medical School, 3311 Yakushiji, Minamikawachi-machi, Tochigi 329-0498, Japan. E-mail: ttakizawa@jichi.ac.jp

We applied a fluorescent ultrasmall immunogold probe, FluoroNanogold (FNG), to immunocytochemistry on ultrathin cryosections. FNG has the properties of both a fluorescent dye-conjugated antibody for fluorescence microscopy and a gold particle-conjugated antibody for electron microscopy. Therefore, this bifunctional immunoprobe permits correlative microscopic observation of the same cell profiles labeled in a single labeling procedure by these two imaging methods. We demonstrate the utility of FNG as a secondary antibody for immunocytochemical labeling of myeloperoxidase (a marker protein for azurophilic granules) in ultrathin cryosectioned human neutrophils. Its detection requires high spatial resolution because neutrophils contain many cytoplasmic granules. There was a one-to-one relationship between fluorescent structures labeled with FNG and organelle profiles labeled with the same silver-enhanced FNG in ultrathin cryosections. Use of FNG immunocytochemistry on ultrathin cryosections is an ideal methodology for high-resolution correlative fluorescence and electron microscopy and can provide unique information that may be difficult to obtain with a single imaging regimen. (J Histochem Cytochem 48:481–485, 2000)

Key Words: FluoroNanogold, immunocytochemistry, correlative microscopy, fluorescence microscopy, electron microscopy, ultrathin cryosections, myeloperoxidase, human neutrophils


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