Patent ID: 6759206
Filing Date: 2004-07-06
Classification: B01J,B01L,B82Y,C12Q,C40B,G01N,Y10S

Abstract:
An automated method for identifying compounds that modulate internalization of cell surface receptor proteins comprising:providing an array of locations containing multiple cells to be treated with a test compound, wherein the cells possess at least a first luminescently labeled reporter molecule that reports on a cell surface receptor protein of interest and at least a second luminescently labeled reporter molecule that identifies cells, and wherein the cell surface receptor protein is either expressed as a luminescently labeled protein, or is luminescently labeled by contacting the cell with a luminescently labeled molecule that binds to the cell surface receptor protein, wherein the contacting can be carried out either before or after treatment with the test compound; contacting the cells with the at least a second luminescently labeled reporter molecule, wherein the contacting can be carried out either before or after treatment with the test compound; treating the cells with the test compound; imaging multiple cells in the array of locations; automatically obtaining luminescent signals from the at least second luminescently labeled reporter molecule to identify cells; automatically obtaining luminescent signals from the at least first luminescently-labeled reporter molecule on or in the cells, comprising: a) acquiring an object image of the at least first luminescently labeled reporter molecule on or in the individual cells; b) by segmenting the object image to produce a segmented object image; and c) calculating whether objects in the segmented object image meet or surpass a user-defined threshold luminescent intensity value, wherein an object that meets or surpasses a user-defined threshold luminescent intensity value represents an internalized cell surface receptor protein of interest: converting the luminescent signals into digital data; and automatically calculating from the digital data: i) a total number of cells, calculated from the digital data from the at least second luminescently labeled reporter molecule; and ii) a number and/or percent of cells that internalized the at least first luminescently labeled reporter molecule; wherein the calculations provide a measure of whether the test compound has modulated internalization of the luminescently labeled cell surface receptor protein.