US5989835A
Expired / Lapsed
US
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System for cell-based screening
Assignee: Cellomics Inc
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Filing: 1997-02-27
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Granted: 1999-11-23
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Expires: 2017-02-27
Abstract
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.
Inventors
- 👤 R. Terry Dunlay
- 👤 D. Lansing Taylor
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 08:21
CPC Classifications
B; B82; B82Y; B82Y30/00; C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; G; G01; G01N; G01N15/00; G01N15/10; G01N15/14; G01N15/1429; G01N15/1433; G01N21/00; G01N21/62; G01N21/63; G01N21/64; G01N21/6428; G01N21/645; G01N21/6452; G01N21/6456; G01N21/6458; G01N33/00; G01N33/48; G01N33/50; G01N33/5005; G01N33/5008; G01N33/502; G01N33/5035; G01N33/58; G01N33/582; G02; G02B; G02B21/00; G02B21/16; G02B21/36; G02B21/365; B01; B01J; B01J2219/00; B01J2219/00274; B01J2219/00277; B01J2219/00279; B01J2219/00306; B01J2219/00313; B01J2219/00315; B01J2219/00317; B01J2219/0068; B01J2219/00702; B01J2219/00707; B01J2219/00718; B01J2219/0072; B01J2219/0074; B01J2219/00743; B01L; B01L3/00; B01L3/50; B01L3/508; B01L3/5085; C40; C40B; C40B60/00; C40B60/14; G01N2015/1486; G01N2015/1497; G01N2021/6417; G01N2021/6419; G01N2021/6482; Y; Y10; Y10S; Y10S436/00; Y10S436/80; Y10S436/807; Y10S436/809; Y10S977/00; Y10S977/70; Y10S977/788; Y10S977/795; Y10S977/84; Y10S977/88; Y10S977/881; Y10S977/902; Y10S977/904; Y10S977/92; Y10S977/932; Y10S977/94; Y10S977/941; Y10S977/942
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
160 words
An automated method for analyzing cells comprising; a) providing an array of locations which contain multiple cells, wherein the cells contain one or more fluorescent reporter molecules; b) scanning multiple cells in each of the locations containing cells to obtain fluorescent signals from the fluorescent reporter molecules in subcellular compartments of the cells being analyzed wherein the subcellular compartments comprise the cell nucleus and the cell cytoplasm; c) converting the fluorescent signals into digital data; d) utilizing the digital data to automatically make measurements, wherein the measurements are used to automatically calculate changes in the distribution, environment or activity of the fluorescent reporter molecules within or between the cell nucleus and the cell cytoplasm of the cells being analyzed, wherein the measurements comprise determining: i) the fluorescence intensity of reporter molecules within the cell nucleus; ii) the fluorescence intensity of reporter molecules within the cell cytoplasm; iii) the area of the cell nucleus; and iv) the area of the cytoplasm.
Claim 2
Dependent Claim
160 words
An automated method for analyzing cells comprising; a) providing an array of locations which contain multiple cells, wherein the cells contain one or more fluorescent reporter molecules; b) scanning multiple cells in each of the locations containing cells to obtain fluorescent signals from the fluorescent reporter molecules in subcellular compartments of the cells being analyzed wherein the subcellular compartments comprise the cell nucleus and the cell cytoplasm; c) converting the fluorescent signals into digital data; d) utilizing the digital data to automatically make measurements, wherein the measurements are used to automatically calculate changes in the distribution, environment or activity of the fluorescent reporter molecules within or between the cell nucleus and the cell cytoplasm of the cells being analyzed, wherein the measurements comprise determining: i) the fluorescence intensity of reporter molecules within the cell nucleus; ii) the fluorescence intensity of reporter molecules within the cell cytoplasm; iii) the area of the cell nucleus; and iv) the area of the cytoplasm.
Claim 3
Dependent Claim
97 words
The method of claim 1 wherein the calculated changes comprise: a) changes in the fluorescence intensity of reporter molecules within the cell nucleus of the cells being analyzed; b) changes in the fluorescence intensity of reporter molecules within the cell cytoplasm of the cells being analyzed; c) changes in the ratio of the fluorescence intensity within the cell cytoplasm to the fluorescence intensity within the cell nucleus of the cells being analyzed; and d) changes in the difference of the fluorescence intensity within the cytoplasm and the fluorescence intensity within the nucleus of the cells being analyzed.
Claim 4
Dependent Claim
29 words
The method of claim 2 wherein the calculated changes provide a measurement of translocation of fluorescent reporter molecules from the cytoplasm to the nucleus of the cells being analyzed.
Claim 5
Dependent Claim
14 words
The method of claim 3 wherein the fluorescent reporter molecules comprise a transcription factor.
Claim 6
Dependent Claim
11 words
The method of claim 4 wherein the transcription factor comprises NF-kB.
Claim 7
Dependent Claim
227 words
An automated method for analyzing cells comprising: a) providing an array of locations which contain multiple cells, wherein the cells contain one or more fluorescent reporter molecules; b) scanning multiple cells in each of the locations containing cells to obtain fluorescent signals from the fluorescent reporter molecules in subcellular compartments of the cells, wherein the subcellular compartments comprise the cell nucleus and a subcellular compartment selected from the group consisting of the cell membrane and the cell cytoplasm; c) converting the fluorescent signals into digital data; d) utilizing the digital data to automatically make measurements, wherein the measurements are used to automatically calculate changes in the distribution, environment or activity of the fluorescent reporter molecules within or between the cell nucleus and the subcellular compartment selected from the group consisting of the cell membrane and the cell cytoplasm, wherein the digital data are captured as pixels, and wherein: i) the fluorescence intensity of reporter molecules within the cell nucleus defines a nuclear region, and wherein the total number of pixels within the nuclear region comprises a measure of nuclear area; and ii) the fluorescence intensity of reporter molecules in the subcellular compartment selected from the group consisting of the cell membrane and the cell cytoplasm defines a cytoplasmic region, and wherein the total number of pixels in the cytoplasmic region comprises a measure of the cytoplasmic area.
Claim 8
Dependent Claim
227 words
An automated method for analyzing cells comprising: a) providing an array of locations which contain multiple cells, wherein the cells contain one or more fluorescent reporter molecules; b) scanning multiple cells in each of the locations containing cells to obtain fluorescent signals from the fluorescent reporter molecules in subcellular compartments of the cells, wherein the subcellular compartments comprise the cell nucleus and a subcellular compartment selected from the group consisting of the cell membrane and the cell cytoplasm; c) converting the fluorescent signals into digital data; d) utilizing the digital data to automatically make measurements, wherein the measurements are used to automatically calculate changes in the distribution, environment or activity of the fluorescent reporter molecules within or between the cell nucleus and the subcellular compartment selected from the group consisting of the cell membrane and the cell cytoplasm, wherein the digital data are captured as pixels, and wherein: i) the fluorescence intensity of reporter molecules within the cell nucleus defines a nuclear region, and wherein the total number of pixels within the nuclear region comprises a measure of nuclear area; and ii) the fluorescence intensity of reporter molecules in the subcellular compartment selected from the group consisting of the cell membrane and the cell cytoplasm defines a cytoplasmic region, and wherein the total number of pixels in the cytoplasmic region comprises a measure of the cytoplasmic area.
Claim 9
Dependent Claim
38 words
The method of claim 6 wherein the calculated changes comprise: (a) changes in the measure of nuclear area; (b) changes in the measure of cytoplasmic area; and (c) changes in the ratio of cytoplasmic area to nuclear area.
Claim 10
Dependent Claim
15 words
The method of claim 7 wherein the calculated changes provide a measurement of cellular hypertrophy.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2002-11-22 | 2003-05-24 | 2003-11-23 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2006-11-22 | 2007-05-24 | 2007-11-23 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2010-11-22 | 2011-05-24 | 2011-11-23 | $7700.00 | Window Closed / Lapsed |