System for cell-based screening
DCFirst Claim
1. An automated method for analyzing translocation of a protein of interest between nucleus and cytoplasm in cells comprising:
- a) providing an array of locations which contain multiple cells, wherein the cells contain at least a first fluorescent reporter molecule that reports on a protein of interest;
b) contacting the cells with at least one test compound of interest;
c) scanning multiple cells in each of the locations containing cells to obtain fluorescent signals from the at least first fluorescent reporter molecule within the nucleus and the cytoplasm in the cells;
d) converting the fluorescent signals into digital data;
e) measuring a total or average fluorescence intensity of the fluorescent signals from the at least first fluorescent reporter molecule within the nucleus and within the cytoplasm of the cells; and
f) automatically calculating test compound-induced changes in one or more of the following;
i) a ratio of fluorescent signal intensity from the at least first fluorescent reporter molecule in the cytoplasm to fluorescent signal intensity from the at least first fluorescent reporter molecule in the nucleus; and
ii) a difference between fluorescent signal intensity from the at least first fluorescent reporter molecule in the cytoplasm and fluorescent signal intensity from the at least first fluorescent reporter molecule in the nucleus;
wherein the test compound-induced changes indicate an effect of the test compound on translocation of the protein of interest between the nucleus and the cytoplasm in the cells.
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Abstract
The invention relates to an optical system for determining the distribution, environment, or activity of fluorescently labeled reporter molecules in cells for the purpose of screening large numbers of compounds for specific biological activity. The invention involves providing cells containing fluorescent reporter molecules in an array of locations and scanning numerous cells in each location with a fluorescent microscope, converting the optical information into digital data, and utilizing the digital data to determine the distribution, environment or activity of the fluorescently labeled reporter molecules in the cells. The array of locations may be an industry standard 96 well or 384 well microtiter plate or a microplate which is a microplate having a cells in a micropaterned array of locations. The invention includes apparatus and computerized method for processing, displaying and storing the data.
130 Citations
15 Claims
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1. An automated method for analyzing translocation of a protein of interest between nucleus and cytoplasm in cells comprising:
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a) providing an array of locations which contain multiple cells, wherein the cells contain at least a first fluorescent reporter molecule that reports on a protein of interest;
b) contacting the cells with at least one test compound of interest;
c) scanning multiple cells in each of the locations containing cells to obtain fluorescent signals from the at least first fluorescent reporter molecule within the nucleus and the cytoplasm in the cells;
d) converting the fluorescent signals into digital data;
e) measuring a total or average fluorescence intensity of the fluorescent signals from the at least first fluorescent reporter molecule within the nucleus and within the cytoplasm of the cells; and
f) automatically calculating test compound-induced changes in one or more of the following;
i) a ratio of fluorescent signal intensity from the at least first fluorescent reporter molecule in the cytoplasm to fluorescent signal intensity from the at least first fluorescent reporter molecule in the nucleus; and
ii) a difference between fluorescent signal intensity from the at least first fluorescent reporter molecule in the cytoplasm and fluorescent signal intensity from the at least first fluorescent reporter molecule in the nucleus;
wherein the test compound-induced changes indicate an effect of the test compound on translocation of the protein of interest between the nucleus and the cytoplasm in the cells. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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Specification