US6900030B2
Expired / Lapsed
US
⏱️ 0.0 years remaining
Device for monitoring cells
Assignee: Becton Dickinson and Co
•
Filing: 2002-04-04
•
Granted: 2005-05-31
•
Expires: 2022-04-04
Abstract
Abstract The present invention relates to methods for detection and evaluation of metabolic activity of eukaryotic and/or prokaryotic cells based upon their ability to consume dissolved oxygen. The methods utilize a luminescence detection system which makes use of the sensitivity of the luminescent emission of certain compounds to the presence of oxygen, which quenches (diminishes) the compound's luminescent emission in a concentration dependent manner. Respiring eukaryotic and/or prokaryotic cells will affect the oxygen concentration of a liquid medium in which they are immersed. Thus, this invention provides a convenient system to gather information on the presence, identification, quantification and cytotoxic activity of eukaryotic and/or prokaryotic cells by determining their effect on the oxygen concentration of the media in which they are present.
Inventors
- 👤 J. Bruce Pitner
- 👤 John Jacob Hemperly
- 👤 Richard D. Guarino
- 👤 Magdalena Wodnicka
- 👤 David T. Stitt
- 👤 Gregory J. Burrell
- 👤 Timothy G. Foley, Jr.
- 👤 Patrick Shawn Beaty
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 05:57
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/02; C12Q1/18; C12Q1/04; C12Q2304/00; C12Q2304/40; C12Q2304/44; Y; Y02; Y02A; Y02A90/00; Y02A90/10
Claims Analysis
Total Claims: 36
Claim 1
Independent Claim
207 words
A method for determining the presence or absence of respiring eukaryotic cells in a solution comprising: (i) contacting said solution with a sensor composition which comprises a luminescent compound that exhibits a change in luminescent property, when irradiated with light containing wavelengths which cause said compound to luminesce, upon exposure to oxygen, wherein the presence of the sensor composition is non-destructive to the eukaryotic cells; (ii) irradiating said sensor composition with light containing wavelengths which cause said luminescent compound to luminesce; (iii) measuring or visually observing the luminescent light intensity from said luminescent compound while irradiating said sensor compound with said light and obtaining a measurement of the luminescence at timed intervals; (iv) comparing said measurement to that of a control not containing respiring eukaryotic cells, wherein said control is selected from the group consisting of: a reagent control not in contact with respiring eukaryotic cells and a calculated threshold, wherein a change in luminescent property relative to the luminescent property of the control is indicative of the presence of respiring eukaryotic cells; and (v) in the event that no such change is measured or observed, repeating steps (ii), (iii), and (iv) to attempt to determine the presence or absence of respiring eukaryotic cells in said solution.
Claim 2
Independent Claim
207 words
A method for determining the presence or absence of respiring eukaryotic cells in a solution comprising: (i) contacting said solution with a sensor composition which comprises a luminescent compound that exhibits a change in luminescent property, when irradiated with light containing wavelengths which cause said compound to luminesce, upon exposure to oxygen, wherein the presence of the sensor composition is non-destructive to the eukaryotic cells; (ii) irradiating said sensor composition with light containing wavelengths which cause said luminescent compound to luminesce; (iii) measuring or visually observing the luminescent light intensity from said luminescent compound while irradiating said sensor compound with said light and obtaining a measurement of the luminescence at timed intervals; (iv) comparing said measurement to that of a control not containing respiring eukaryotic cells, wherein said control is selected from the group consisting of: a reagent control not in contact with respiring eukaryotic cells and a calculated threshold, wherein a change in luminescent property relative to the luminescent property of the control is indicative of the presence of respiring eukaryotic cells; and (v) in the event that no such change is measured or observed, repeating steps (ii), (iii), and (iv) to attempt to determine the presence or absence of respiring eukaryotic cells in said solution.
Claim 3
Dependent Claim
31 words
The method of claim 1 wherein said luminescent compound is contained within a matrix which is relatively impermeable to water and non-gaseous solutes, but which has a high permeability to oxygen.
Claim 4
Dependent Claim
14 words
The method of claim 2 wherein said matrix is a rubber or plastic matrix.
Claim 5
Dependent Claim
13 words
The method of claim 2 wherein said matrix is a silicone rubber matrix.
Claim 6
Dependent Claim
15 words
The method of claim 2 wherein said luminescent compound is adsorbed on solid silica particles.
Claim 7
Dependent Claim
15 words
The method of claim 1 wherein said luminescent compound is a tris-4,7-diphenyl-1,10-phenanthroline ruthenium (II) salt.
Claim 8
Dependent Claim
14 words
The method of claim 6 wherein said luminescent compound is tris-4,7-diphenyl-1,10-phenanthroline ruthenium (II) chloride.
Claim 9
Dependent Claim
15 words
The method of claim 1 wherein said luminescent compound is a tris-2,2′-bipyridyl ruthenium (II) salt.
Claim 10
Dependent Claim
15 words
The method of claim 8 wherein said luminescent compound is tris-2,2′-bipyridyl ruthenium (II) chloride hexahydrate.
Claim 11
Dependent Claim
12 words
The method of claim 1 wherein said luminescent compound is 9,10-diphenyl anthracene.
Claim 12
Dependent Claim
22 words
The method of claim 1 wherein said solution is isolated from atmospheric oxygen wherein said solution is contained in a closed system.
Claim 13
Dependent Claim
13 words
The method of claim 1 wherein said solution is exposed to atmospheric oxygen.
Claim 14
Dependent Claim
58 words
The method of claim 1 wherein, in step (i), the solution is also contacted with an effective concentration of one or more biomaterials, selected from the group consisting of laminin, collagen IV, entactin, heparin sulfate proteoglycans, defined mammalian and insect cell growth factors, and matrix metalloproteinases, said concentration being effective for promoting or enabling cellular growth and respiration.
Claim 15
Dependent Claim
20 words
The method of claim 13 wherein said one or more biomaterials are laminin, collagen IV, entactin and heparin sulfate proteoglycans.
Claim 16
Dependent Claim
36 words
The method of claim 1 wherein, in step (i), the solution is also contacted with an effective concentration of one or more extracellular matrices, said concentration being effective for promoting or enabling cellular growth and respiration.
Claim 17
Dependent Claim
11 words
The method of claim 15 wherein said extracellular matrix is collagen.
Claim 18
Dependent Claim
55 words
The method of claim 1 wherein, in step (i), the solution is contacted with an effective concentration of one or more additives or coating substances, selected from the group consisting of penicillin, streptomycin, fungizone, non-essential amino acids, sodium pyruvate, and fetal bovine serum, said concentration being effective for promoting or enabling cellular growth and respiration.
Claim 19
Independent Claim
198 words
A method for quantifying respiring eukaryotic cells in a solution comprising: (i) contacting said solution with a sensor composition which comprises a luminescent compound that exhibits a change in luminescent property, when irradiated with light containing wavelengths which cause said compound to luminesce, upon exposure to oxygen, wherein the presence of the sensor composition is non-destructive to the eukaryotic cells; (ii) irradiating said sensor composition with light containing wavelengths which cause said luminescent compound to luminesce; (iii) measuring or visually observing the change iii luminescent property from said luminescent compound while irradiating said sensor compound with said light and obtaining a measurement of the luminescence at timed intervals; (iv) comparing said measurement to that of a control not containing respiring eukaryotic cells, wherein said control is selected from the group consisting of: a reagent control not in contact with respiring eukaryotic cells and a calculated threshold, wherein a change in luminescent property relative to the luminescent property of the control is indicative of the presence of respiring eukaryotic cells; and (v) in the event that no such change is measured or observed, repeating steps (ii), (iii), and (iv), to attempt to quantify respiring eukaryotic cells in said solution.
Claim 20
Independent Claim
198 words
A method for quantifying respiring eukaryotic cells in a solution comprising: (i) contacting said solution with a sensor composition which comprises a luminescent compound that exhibits a change in luminescent property, when irradiated with light containing wavelengths which cause said compound to luminesce, upon exposure to oxygen, wherein the presence of the sensor composition is non-destructive to the eukaryotic cells; (ii) irradiating said sensor composition with light containing wavelengths which cause said luminescent compound to luminesce; (iii) measuring or visually observing the change iii luminescent property from said luminescent compound while irradiating said sensor compound with said light and obtaining a measurement of the luminescence at timed intervals; (iv) comparing said measurement to that of a control not containing respiring eukaryotic cells, wherein said control is selected from the group consisting of: a reagent control not in contact with respiring eukaryotic cells and a calculated threshold, wherein a change in luminescent property relative to the luminescent property of the control is indicative of the presence of respiring eukaryotic cells; and (v) in the event that no such change is measured or observed, repeating steps (ii), (iii), and (iv), to attempt to quantify respiring eukaryotic cells in said solution.
Claim 21
Dependent Claim
31 words
The method of claim 18 wherein said luminescent compound is contained within a matrix which is relatively impermeable to water and non-gaseous solutes, but which has a high permeability to oxygen.
Claim 22
Dependent Claim
14 words
The method of claim 19 wherein said matrix is a rubber or plastic matrix.
Claim 23
Dependent Claim
13 words
The method of claim 19 wherein said matrix is a silicone rubber matrix.
Claim 24
Dependent Claim
15 words
The method of claim 19 wherein said luminescent compound is adsorbed on solid silica particles.
Claim 25
Dependent Claim
15 words
The method of claim 18 wherein said luminescent compound is a tris-4,7-diphenyl-1,10-phenanthroline ruthenium (ii) salt.
Claim 26
Dependent Claim
14 words
The method of claim 23 wherein said luminescent compound is tris4,7-diphenyl-1,10-phenanthroline ruthenium (II) chloride.
Claim 27
Dependent Claim
15 words
The method of claim 18 wherein said luminescent compound is a tris-2,2′-bipyridyl ruthenium (II) salt.
Claim 28
Dependent Claim
15 words
The method of claim 25 wherein said luminescent compound is tris-2,2′-bipyridyl ruthenium (II) chloride hexahydrate.
Claim 29
Dependent Claim
12 words
The method of claim 18 wherein said luminescent compound is 9,10-diphenyl anthracene.
Claim 30
Dependent Claim
22 words
The method of claim 18 wherein said solution is isolated from atmospheric oxygen wherein said solution is contained in a closed system.
Claim 31
Dependent Claim
13 words
The method of claim 18 wherein said solution is exposed to atmospheric oxygen.
Claim 32
Dependent Claim
58 words
The method of claim 18 wherein, in step (i), the solution is also contacted with an effective concentration of one or more biomaterials, selected from the group consisting of laminin, collagen IV, entactin, heparin sulfate proteoglycans, defined mammalian and insect cell growth factors, and matrix metalloproteinases, said concentration being effective for promoting or enabling cellular growth and respiration.
Claim 33
Dependent Claim
20 words
The method of claim 30 wherein said one or more biomaterials are laminin, collagen IV, entactin and heparin sulfate proteoglycans.
Claim 34
Dependent Claim
36 words
The method of claim 18 wherein, in step (i), the solution is also contacted with an effective concentration of one or more extracellular matrices, said concentration being effective for promoting or enabling cellular growth and respiration.
Claim 35
Dependent Claim
11 words
The method of claim 32 wherein said extracellular matrix is collagen.
Claim 36
Dependent Claim
55 words
The method of claim 18 wherein, in step (i), the solution is contacted with an effective concentration of one or more additives or coating substances selected from the group consisting of penicillin, streptomycin, fungizone, non-essential amino acids, sodium pyruvate, and fetal bovine serum, said concentration being effective for promoting or enabling cellular growth and respiration.
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