Patent ID: 6620591
Filing Date: 2003-09-16
Classification: B01J,B01L,B82Y,C12Q,C40B,G01N,G02B,Y10S

Abstract:
An automated method for detecting translocation of a cellular component of interest between a first cellular compartment and a second cellular compartment on and/or within individual cells comprising:a) providing an array of locations which contain multiple cells, wherein the cells contain at least a first fluorescent reporter molecule to identify individual cells, and at least a second fluorescent reporter molecule to report on a cellular component of interest, wherein fluorescent signals from the at least first and the at least second fluorescent reporter molecules are optically distinguishable; b) automatically scanning multiple cells in each of the locations containing cells to obtain fluorescent signals from the at least first fluorescent reporter molecule, wherein the scanning comprises: i) acquiring fluorescent signals in a field from the at least first fluorescent reporter molecule at a first wavelength; ii) automatically converting the fluorescent signals from the at least first fluorescent reporter molecule into digital data; and iii) automatically utilizing the digital data from the at least first fluorescent reporter molecule to identify individual cells in the field; c) automatically acquiring fluorescent signals from the at least second fluorescent reporter molecule at a second wavelength in a field containing individual cells; d) automatically converting the fluorescent signals from the at least second fluorescent reporter molecule into digital data; e) automatically defining a first cellular compartment mask and a second cellular compartment mask from the fluorescent signals from the at least first reporter molecule and the at least second fluorescent reporter molecule; f) automatically determining an intensity of the fluorescent signals from the at least second fluorescent reporter molecule in the first cellular compartment mask and the second cellular compartment mask; and g) automatically calculating one or both of the following: i) a ratio of the intensity of the fluorescent signals from the at least second fluorescent reporter molecule in the first cellular compartment mask and the second cellular compartment mask; and ii) a difference of the intensity of the fluorescent signals from the at least second fluorescent reporter molecule in the first cellular compartment mask and the second cellular compartment mask; wherein the ratio of the intensity of the fluorescent signals from the at least second fluorescent reporter molecule in the first cellular compartment mask and the second cellular compartment mask and/or the difference of the intensity of the fluorescent signals from the at least second fluorescent reporter molecule in the first cellular compartment mask and the second cellular compartment mask provides a measure of the translocation of the cellular component of interest between the first cellular compartment and the second cellular compartment on and/or within the individual cells.