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Molecular Pharmacology, Vol 10, 986-998, Copyright © 1974 by the American Society for Pharmacology and Experimental Therapeutics

A Correlative Radioautographic, Fluorescent, and Histochemical Technique for Cytopharmacology

LLOYD J. ROTH 1, IHSAN M. DIAB 1, MITSUTOSHI WATANABE 1, and ROBERT J. DINERSTEIN 1

1 Department of Pharmacological and Physiological Sciences, The University of Chicago, Chicago, Illinois 60637

Using mouse intestine, a model system has been developed in which two target cell loci, biogenic amine-containing enterochromaffin cells and regenerating crypt cells, are used for correlation of drug localization with fluorescence and microscopic structure. The combined procedure described for the cellular localization of drug and fluorophore is based on the use of frozen freeze-dried sections, dry-mounted on dried photographic emulsion, and has general applicability. Tissues need not be subjectected to contact with any solvents until after radioautography, cellular fluorescence, and microspectrophotofluorometry have been completed. The use of freeze-dried frozen sections provides the possibility for localization by immunofluorescence labeling and thus offers the opportunity, by combining radioautographic and fluorescence labeling, to investigate competitive inhibition at the cellular level. Because this technique provides for reversible separation of the tissue from the emulsion for independent treatment, histochemical methods that are potentially destructive for the radioautogram can also be used. The correlative technique described here is rigorously controlled and systematic, and the results are unambiguous.

Submitted on June 24, 1974




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