Patent ID: 6671624
Filing Date: 2003-12-30
Classification: B01J,B82Y,C12Q,C40B,G01N

Abstract:
A machine readable storage medium comprising a program containing a set of instructions for causing a cell screening system to execute procedures for detecting the distribution of one or more cellular macromolecule of interest between a nucleus and cytoplasm in individual cells, comprising:a) scanning multiple cells in an array of locations which contain multiple cells to obtain fluorescent signals from fluorescent reporter molecules in the cells, wherein the cells possess a plurality of fluorescent reporter molecules, wherein the plurality of fluorescent reporter molecules comprises fluorescent reporter molecules to report on: (i) one or more cellular macromolecule of interest; (ii) the cell nucleus; and (iii) the cell cytoplasm; b) identifying individual cells from the fluorescent signals from the plurality of fluorescent reporter molecules; c) creating a nuclear mask and a cytoplasmic mask in the individual cells from the fluorescent signals from the plurality of fluorescent reporter molecules; d) determining an intensity of the fluorescent signals from the fluorescent reporter molecules that report on the one or more cellular macromolecule of interest within the nuclear mask and within the cytoplasmic mask in the individual cells in response to contacting the cells with a test stimulus; e) calculating one or more of the following: i) a ratio of average intensity of the fluorescent signals from the fluorescent reporter molecules that report on the one or more cellular macromolecule of interest between the nuclear mask and the cytoplasmic mask in the multiple cells; ii) a difference of average intensity of the fluorescent signals from the fluorescent reporter molecules that report on the one or more cellular macromolecule of interest between the nuclear mask and the cytoplasmic mask in the multiple cells; iii) a ratio of integrated intensity of the fluorescent signals from the fluorescent reporter molecules that report on the one or more cellular macromolecule of interest between the nuclear mask and the cytoplasmic mask in the multiple cells; and iv) a difference of integrated intensity of the fluorescent signals from the fluorescent reporter molecules that report on the one or more cellular macromolecule of interest between the nuclear mask and the cytoplasmic mask in the multiple cells; f) comparing the calculations to similar calculations made: i) in the cells in response to contacting the cells with the test stimulus from at least a second time point; and/or ii) in the individual cells that have not been contacted with the test stimulus; and g) determining the effect of the test stimulus on the distribution of the one or more cellular macromolecule of interest between the nucleus and the cytoplasm in the individual cells from the calculations.