System for cell-based screening

  • US 6,573,039 B1
  • Filed: 08/29/2000
  • Issued: 06/03/2003
  • Est. Priority Date: 02/27/1997
  • Status: Expired due to Term
First Claim
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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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