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Journal of Histochemistry and Cytochemistry, Vol. 45, 155-158, Copyright © 1997 by The Histochemical Society, Inc.

RAPID COMMUNICATION

Quadruple Immunofluorescence: A Direct Visualization Method

Gian-Luca Ferria, Rosa M. Gaudioa, Ida F. Castelloa, Peter Bergerb, and Gabriele Giroc
a Neuro- and Endocrine Research, Department of Cytomorphology, University of Cagliari, "Oasi" IRCCS, Troina, Italy
b Institute for Biomedical Aging Research, Austrian Academy of Sciences, Innsbruck, Austria
c FRAE Institute, CNR, Bologna, Italy

Correspondence to: Gian-Luca Ferri, Dept. of Cytomorphology, via Porcell 2, 09124 Cagliari, Italy.

We describe fluorescence immunostaining of four different antigens in the same section. The fluorochrome conjugates used show highest emission in the blue, green, yellow, and red regions of the visible light spectrum, respectively. Specially designed single fluorochrome filter combinations allow selective visualization of each fluorochrome label in turn, without visible crosstalk with the others. (J Histochem Cytochem 45:155-158, 1997)

Key Words: immunohistochemistry, quadruple immunostaining, multiple immunostaining, fluorochromes, interference filters, co-localization


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