Cytological assay procedure

G - Physics – 01 – N

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G01N 33/52 (2006.01) A61B 5/0488 (2006.01) G01N 33/487 (2006.01) G01N 33/50 (2006.01)

Patent

CA 1132890

CYTOLOGICAL ASSAY PROCEDURE ABSTRACT OF THE DISCLOSURE Disclosed is a method of bioassay based on the detec- tion of changes in the membrane potential of individual cells. A cell suspension is incubated with a solution of a substance capable of affecting the physiology of at least a subpopulation of cells in the suspension. During or after the incubation, a measurement representative of the membrane potential of indivi- dual cells is made. Comparison of these measurements with reference measurements provides information concerning: 1) the viability of the cells or a subgroup thereof after exposure to toxic substances; 2) the presence and concentration of factors or ligands in the solution which bind specifically to cell membrane receptors; 3) the presence of cells in the suspension sensitive to factors known to be present in the solution; 4) the cumulative effect of plural factors in the solution on the cells or a sub- population thereof; and 5) comparisons among the effects on the cells of plural, chemically distinct factors. The preferred method of measuring membrane potential involves incubating the cells with a membrane-permeant, fluorescent, ionic dye which becomes distributed on opposite sides of the cell membrane as a function of membrane potential, and then assaying intracellular dye concentration by photometric measurements in a flow cyto- meter. - 1 -

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