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Journal of Histochemistry and Cytochemistry, Vol. 51, 401-404, March 2003, Copyright © 2003, The Histochemical Society, Inc.

BRIEF REPORT

Localization of GFP in Frozen Sections from Unfixed Mouse Tissues: Immobilization of a Highly Soluble Marker Protein by Formaldehyde Vapor

Harald Jockuscha, Sylvana Voigta, and Daniel Eberharda
a Developmental Biology and Molecular Pathology, University of Bielefeld, Bielefeld, Germany

Correspondence to: Harald Jockusch, Developmental Biology and Molecular Pathology, W7, University of Bielefeld, D-33501 Bielefeld, Germany. E-mail: h.jockusch@uni-bielefeld.de

Green fluorescent protein (GFP) and its variants, such as enhanced GFP (EGFP), have been introduced into mammalian cells by transgenes, e.g., to distinguish donor from host cells after transplantation. Free GFP is extremely soluble and leaks out from liquid-covered cryostat sections so that fixation of whole organs before sectioning has been mandatory. This precludes the analysis of serial sections with respect to fixation-sensitive enzyme activities and antigens. We describe here a vapor fixation for sections from unfixed cryostat blocks of tissue that allows unrestricted enzyme and immunohistochemistry on adjacent sections, as demonstrated for cross-striated muscle and other tissues from EGFP transgenic "green mice" and for a transplantation experiment.

(J Histochem Cytochem 51:401–404, 2003)

Key Words: green fluorescent protein, transgenes, vapor fixation


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