Inventors:
Howard M. Shapiro - West Newton MA
International Classification:
G01N 130, G01N 1300, G01N 2125, G01N 3348, G01N 3352, G01N 3354, G01N 3374
Abstract:
Disclosed is a method ofbioassay based on the detection 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 individual 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 subpopulation 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 cytometer.