US6875578B2
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US
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System for cell-based screening
Assignee: Cellomics Inc
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Filing: 2002-03-19
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Granted: 2005-04-05
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Expires: 2022-03-19
Abstract
Abstract The present invention provides systems, methods, screens, reagents and kits for optical system analysis of cells to rapidly determine the distribution, environment, or activity of fluorescently labeled reporter molecules in cells for the purpose of screening large numbers of compounds for those that specifically affect particular biological functions.
Inventors
- 👤 Kenneth Giuliano
- 👤 Ravi Kapur
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 23:30
CPC Classifications
G; G01; G01N; G01N21/00; G01N21/62; G01N21/63; G01N21/64; G01N21/645; G01N21/6456; G01N21/6458; B; B82; B82Y; B82Y30/00; C; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C12N15/1034; C12N5/00; C12N5/0062; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; G01N15/00; G01N15/10; G01N15/14; G01N15/1429; G01N15/1433; G01N21/6428; G01N21/6452; G01N33/00; G01N33/48; G01N33/50; G01N33/5005; G01N33/5008; G01N33/5011; G01N33/5014; G01N33/502; G01N33/5076; G01N33/5079; G01N33/53; G01N33/543; G01N33/54353; G01N33/54366; G01N33/58; G01N33/582; 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; B01L3/502; B01L3/5027; C40; C40B; C40B60/00; C40B60/14; G01N15/1468; G01N2015/1486; G01N2015/1497; Y; Y10; Y10S; Y10S436/00; Y10S436/80; Y10S436/807; Y10S436/808; Y10S436/809
Claims Analysis
Total Claims: 34
Claim 1
Independent Claim
324 words
An automated method for cell based toxin detection, classification, and/or identification comprising providing an array of locations containing cells to be treated with a test substance, wherein the cells possess at least a first fluorescent reporter molecule comprising a detector, wherein the detector detects a toxin present in the test substance, and a second fluorescent reporter molecule selected from the group consisting of a classifier or an identifier, wherein the classifier detects a toxin present in the test substance and identifies a cell pathway affected by a toxin present in the test substance, and the identifier detects a toxin present in the test substance and identifies a specific toxin or group of toxins present in the test substance; contacting the cells with the test substance either before or after possession of the first and second fluorescent reporter molecules by the cells; wherein the localization, distribution, structure, or activity of the first and second fluorescent reporter molecule is modified when the cell is contacted with a toxin present in the test substance; imaging or scanning multiple cells in each of the locations containing cells to obtain fluorescent signals from the detector and the second reporter molecule; converting the fluorescent signals from the detector and the second reporter molecule into digital data; utilizing the digital data from the detector and the second reporter molecule to automatically measure the localization, distribution, or activity of the detector and the second reporter molecule on or in the cell, wherein a change in the localization, distribution, structure or activity of the detector indicates the presence of a toxin in the test substance, and; wherein a change in the localization, distribution, structure or activity of the classifier identifies a cell pathway that is perturbed by the toxin present in the test substance, or wherein a change in the localization, distribution, structure or activity of the identifier identifies the specific toxin or group of toxins that are present in the test substance.
Claim 2
Dependent Claim
324 words
An automated method for cell based toxin detection, classification, and/or identification comprising providing an array of locations containing cells to be treated with a test substance, wherein the cells possess at least a first fluorescent reporter molecule comprising a detector, wherein the detector detects a toxin present in the test substance, and a second fluorescent reporter molecule selected from the group consisting of a classifier or an identifier, wherein the classifier detects a toxin present in the test substance and identifies a cell pathway affected by a toxin present in the test substance, and the identifier detects a toxin present in the test substance and identifies a specific toxin or group of toxins present in the test substance; contacting the cells with the test substance either before or after possession of the first and second fluorescent reporter molecules by the cells; wherein the localization, distribution, structure, or activity of the first and second fluorescent reporter molecule is modified when the cell is contacted with a toxin present in the test substance; imaging or scanning multiple cells in each of the locations containing cells to obtain fluorescent signals from the detector and the second reporter molecule; converting the fluorescent signals from the detector and the second reporter molecule into digital data; utilizing the digital data from the detector and the second reporter molecule to automatically measure the localization, distribution, or activity of the detector and the second reporter molecule on or in the cell, wherein a change in the localization, distribution, structure or activity of the detector indicates the presence of a toxin in the test substance, and; wherein a change in the localization, distribution, structure or activity of the classifier identifies a cell pathway that is perturbed by the toxin present in the test substance, or wherein a change in the localization, distribution, structure or activity of the identifier identifies the specific toxin or group of toxins that are present in the test substance.
Claim 3
Dependent Claim
38 words
The method of claim 1 wherein the second fluorescent reporter molecule is a classifier, and the digital data derived from the classifier is used to select an identifier for identification of the specific toxin or group of toxins.
Claim 4
Dependent Claim
20 words
The method of claim 1 , wherein the first fluorescent reporter and the second fluorescent reporter molecules are different molecules.
Claim 5
Dependent Claim
12 words
The method of claim 3 , wherein the cells are eukaryotic cells.
Claim 6
Dependent Claim
11 words
The method of claim 4 wherein the cells are mammalian cells.
Claim 7
Dependent Claim
48 words
The method of claim 5 wherein the detector comprises a molecule selected from the group consisting of heat shock proteins and compounds that respond to changes in mitochondrial membrane potential, intracellular free ion concentration, cytoskeletal organization, general metabolic status, cell cycle timing events, and organellar structure and function.
Claim 8
Dependent Claim
28 words
The method of claim 5 wherein the classifier comprises a molecule selected from the group consisting of tubulin, microtubule-associated proteins, actin, actin-binding proteins, NF-κB, IκB, and stress-activated kinases.
Claim 9
Dependent Claim
32 words
The method of claim 5 wherein the cell pathway is selected from the group consisting of cell stress pathways, cell metabolic pathways, cell signaling pathways, cell growth pathways, and cell division pathways.
Claim 10
Dependent Claim
39 words
The method of claim 5 , wherein the second fluorescent reporter molecule is an identifier, and the identifier identifies a toxin or group of toxins selected from the group consisting of proteases, ADP-ribosylating toxins, cytotoxic phospholipases, and exfoliative toxins.
Claim 11
Dependent Claim
81 words
The method of claim 5 wherein the change in the localization, distribution, structure or activity of the first and/or second fluorescent reporter molecules is selected from the group consisting of cytoplasm to nucleus translocation, nucleus or nucleolus to cytoplasm translocation, receptor internalization, mitochondrial membrane potential, loss of signal, the spectral response of the reporter molecule, phosphorylation, intracellular free ion concentration, cell size, cell shape, cytoskeleton organization, metabolic processes, cell motility, cell substrate attachment, cell cycle events, and organellar structure and function.
Claim 12
Dependent Claim
15 words
The method of claim 3 , wherein the method is used to identify toxin mimetics.
Claim 13
Dependent Claim
15 words
The method of claim 3 , wherein the method is used to identify toxin inhibitors.
Claim 14
Dependent Claim
14 words
The method of claim 3 , wherein the test substance is an environmental sample.
Claim 15
Dependent Claim
16 words
The method of claim 13 wherein the environmental sample is a development or manufacturing process sample.
Claim 16
Dependent Claim
18 words
The method of claim 13 wherein the environmental sample may comprise toxins utilized in chemical and biological weapons.
Claim 17
Dependent Claim
14 words
The method of claim 13 wherein the environmental sample is an environmental remediation sample.
Claim 18
Dependent Claim
42 words
The method of claim 3 wherein the test substance is selected from the group consisting of a control sample known to contain a toxin, a control sample known to not contain a toxin, and a test sample with an unknown toxin content.
Claim 19
Dependent Claim
144 words
The method of claim 3 , wherein the second fluorescent reporter molecule comprises a classifier, and wherein the cells further possess at least a third fluorescent reporter molecule comprising an identifier; wherein the contacting the cells with the test substance is done either before or after possession of the third fluorescent reporter molecules by the cells; wherein the localization, distribution, structure, or activity of the third fluorescent reporter molecules is modified when the cell is contacted with the toxin, and wherein the method further comprises: imaging or scanning multiple cells in each of the locations containing multiple cells to obtain fluorescent signals from the identifier; converting the fluorescent signals from the identifier into digital data; utilizing the digital data from the third fluorescent reporter molecule to automatically measure the localization, distribution, or activity of the third fluorescent reporter molecule on or in the cell.
Claim 20
Dependent Claim
28 words
The method of claim 18 , wherein each of the first fluorescent reporter molecule, the second fluorescent reporter molecule, and the third fluorescent reporter molecule are different molecules.
Claim 21
Dependent Claim
12 words
The method of claim 19 , wherein the cells are eukaryotic cells.
Claim 22
Dependent Claim
11 words
The method of claim 20 wherein the cells are mammalian cells.
Claim 23
Dependent Claim
48 words
The method of claim 21 wherein the detector comprises a molecule selected from the group consisting of heat shock proteins and compounds that respond to changes in mitochondrial membrane potential, intracellular free ion concentration, cytoskeletal organization, general metabolic status, cell cycle timing events, and organellar structure and function.
Claim 24
Dependent Claim
28 words
The method of claim 21 wherein the classifier comprises a molecule selected from the group consisting of tubulin, microtubule-associated proteins, actin, actin-binding proteins, NE-κB, IκB, and stress-activated kinases.
Claim 25
Dependent Claim
32 words
The method of claim 21 wherein the cell pathway is selected from the group consisting of cell stress pathways, cell metabolic pathways, cell signaling pathways, cell growth pathways, and cell division pathways.
Claim 26
Dependent Claim
30 words
The method of claim 21 , wherein the identifier identifies a toxin or group of toxins selected from the group consisting of proteases, ADP-ribosylating toxins, cytotoxic phospholipases, and exfoliative toxins.
Claim 27
Dependent Claim
82 words
The method of claim 21 wherein the change in the localization, distribution, structure or activity of the first, second, and/or third fluorescent reporter molecules is selected from the group consisting of cytoplasm to nucleus translocation, nucleus or nucleolus to cytoplasm translocation, receptor internalization, mitochondrial membrane potential, loss of signal, the spectral response of the reporter molecule, phosphorylation, intracellular free ion concentration, cell size, cell shape, cytoskeleton organization, metabolic processes, cell motility, cell substrate attachment, cell cycle events, and organellar structure and function.
Claim 28
Dependent Claim
15 words
The method of claim 18 , wherein the method is used to identify toxin mimetics.
Claim 29
Dependent Claim
15 words
The method of claim 18 , wherein the method is used to identify toxin inhibitors.
Claim 30
Dependent Claim
14 words
The method of claim 18 , wherein the test substance is an environmental sample.
Claim 31
Dependent Claim
16 words
The method of claim 29 wherein the environmental sample is a development or manufacturing process sample.
Claim 32
Dependent Claim
18 words
The method of claim 29 wherein the environmental sample may comprise toxins utilized in chemical and biological weapons.
Claim 33
Dependent Claim
14 words
The method of claim 29 wherein the environmental sample is an environmental remediation sample.
Claim 34
Dependent Claim
42 words
The method of claim 29 wherein the test substance is selected from the group consisting of a control sample known to contain a toxin, a control sample known to not contain a toxin, and a test sample with an unknown toxin content.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 2008-04-04 | 2008-10-04 | 2009-04-05 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2012-04-04 | 2012-10-04 | 2013-04-05 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2016-04-04 | 2016-10-04 | 2017-04-05 | $7700.00 | Window Closed / Lapsed |