Fluorochrome Labeling

Richard R Hardy, Fox Chase Cancer Center, Philadelphia, USA

Fluorochrome-labeled reagents are an important tool in the analysis of cell surface antigens, both for use in fluorescence microscopy and in flow cytometry. The development of the hybridoma technique for the generation of monoclonal antibodies that can then be employed as fluorescent reagents has greatly aided the delineations of numerous functional subpopulations now known to constitute the immune system.

Furthermore, the workhorses of microscopy, fluorescein isothiocyanate (FITC, fluorescing green) and tetramethyl rhodamine isothiocyanate (TRITC, fluorescing red) have been joined in recent years by new dyes, the photosynthetic accessory proteins known as phycobiliproteins, possessing intense fluorescence and useful excitation/emission spectra. The techniques used for generating fluorescent reagents using small organic dyes are relatively simple, while those employed for coupling large proteins such as phycobiliproteins are more involved.

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