©J.Paul Robinson, Purdue University BMS 631 - LECTURE1.PPT
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Fluorescence
Labeling Technique
Coons et al
1941 - developed the fluorescence
antibody technique - they labeled antipneumococcal antibodies with anthracine allowing them to detect both the organism and the antibody in tissue using UV
excited blue fluorescence
Manuscript:
Immunological
Properties of an Antibody Containing a Fluorescent Group
Albert
H. Coons, Hugh J. Creech and R. Norman Jones
Department of
Bacteriology and Immunology, Harvard Medical School, and the Chemical Laboratory, Harvard
University
Proc. Soc.
Exp.Biol.Med. 47:200-202, 1941
“Moreover, when Type II and III organisms were dried on different parts of the same slide, exposed to the conjugate for 30 minutes, washed in saline and distilled water, and mounted in glycerol, individual Type III organisms could be seen with the fluorescence microscope……”
Coons and Kaplan (1950) - conjugated fluorescein with isocyanate - better blue green fluorescent signal - further away from tissue autofluorescence. This method used a very
dangerous preparative step using
phosgene gas
