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Heat-Induced Antigen Retrieval Applied in Zebrafish: Whole-Mount In Situ Immunofluorescence Microscopy

Published online by Cambridge University Press:  12 April 2012

Chuang-yu Lin
Department of Chemical Engineering, National Taipei University of Technology, Taipei 106, Taiwan
Wen-ta Su
Department of Chemical Engineering, National Taipei University of Technology, Taipei 106, Taiwan
Li-tzu Li*
Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan
Corresponding author. E-mail:
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Whole-mount immunofluorescence technique provides a way to reveal integrated expression patterns of biological molecules in individuals. Well-documented morphological preservation ability in biology makes aldehydes the fixative of choice. Cross-linking among biocomponents and aldehydes is the key for maintaining morphology but masks the biological molecules for immunodetection. This study performs an easily accessible method by applying heat-induced retrieval, which can rescue the antigenicity of the proteins and also enhance the labeling sensitivity of the fluorescence dye in overfixed zebrafish embryos. The results show that the immunoreactivities of antibodies to myosin in the muscles, green fluorescent protein in the blood vessels and the nuclei in the cells can be recovered significantly, and the morphology of the zebrafish embryos, even the fragile mutants, is at the same time well maintained. Therefore, we provide a choice for antigen retrieval, which is effective for whole-mount immunofluorescence microscopy.

Biological Applications
Copyright © Microscopy Society of America 2012

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