Device for monitoring cells
First Claim
1. 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.
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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.
37 Citations
34 Claims
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1. A method for determining the presence or absence of respiring eukaryotic cells in a solution comprising:
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(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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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18. A method for quantifying respiring eukaryotic cells in a solution comprising:
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(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. - View Dependent Claims (19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34)
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Specification