Metabolic rate indicator for cellular populations
First Claim
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1. A method of determining a metabolic rate indicator of a cellular population, the method comprising:
- combining a sample containing a cellular population and a bioreporter selected from di-O-propionylfluorescein, di-O-acetylfluorescein, O-acetylfluorescein, 6,8-difluoro-4-methylumbelliferyl butyrate, and resorufin acetate, and optionally an additive, in a vessel equipped with a parameter-detecting device for detecting a parameter, the parameter comprising weight of the sample, volume of the sample, or a combination thereof, wherein combining the sample with the bioreporter and/or the optional additive causes a change in the detected parameter and constitutes an initial point of reaction;
detecting temperature, turbidity, and metabolic signal of the sample at the initial point of reaction, wherein the metabolic signal correlates to conversion of the bioreporter by the cellular population to a molecule capable of generating a detectable fluorescent signal;
detecting the temperature, the turbidity, and the metabolic signal of the sample at one or more points in time after the initial point of reaction;
normalizing the detected metabolic signals for differences in detected temperature by using a temperature normalization algorithm and detected turbidity by calibrating against a mixture of the sample and a standard solution at the initial point of reaction and the one or more points in tune after the initial point of reaction, thereby providing a normalized metabolic signal for the initial point of reaction and for each of the one or more points in time; and
determining the metabolic rate indicator of the sample based upon the normalized metabolic signals, wherein rate of conversion of the bioreporter by the cellular population to the molecule capable of generating the detectable fluorescent signal correlates with rate of carbon dioxide production or rate of ethanol production by the cellular population.
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Abstract
Described and claimed is a novel system and method for monitoring the metabolic rate of a cellular population in an industrial process. The invention includes obtaining a sample of the cellular population and initiating a reaction between a metabolic component of the cellular population and a bioreporter. The reaction produces a detectable metabolic signal which is measured and analyzed to determine a metabolic rate of the cellular population. In preferred embodiments, a controller is operable to provide prompts to a user and/or automate various steps of the invention.
18 Citations
12 Claims
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1. A method of determining a metabolic rate indicator of a cellular population, the method comprising:
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combining a sample containing a cellular population and a bioreporter selected from di-O-propionylfluorescein, di-O-acetylfluorescein, O-acetylfluorescein, 6,8-difluoro-4-methylumbelliferyl butyrate, and resorufin acetate, and optionally an additive, in a vessel equipped with a parameter-detecting device for detecting a parameter, the parameter comprising weight of the sample, volume of the sample, or a combination thereof, wherein combining the sample with the bioreporter and/or the optional additive causes a change in the detected parameter and constitutes an initial point of reaction; detecting temperature, turbidity, and metabolic signal of the sample at the initial point of reaction, wherein the metabolic signal correlates to conversion of the bioreporter by the cellular population to a molecule capable of generating a detectable fluorescent signal; detecting the temperature, the turbidity, and the metabolic signal of the sample at one or more points in time after the initial point of reaction; normalizing the detected metabolic signals for differences in detected temperature by using a temperature normalization algorithm and detected turbidity by calibrating against a mixture of the sample and a standard solution at the initial point of reaction and the one or more points in tune after the initial point of reaction, thereby providing a normalized metabolic signal for the initial point of reaction and for each of the one or more points in time; and determining the metabolic rate indicator of the sample based upon the normalized metabolic signals, wherein rate of conversion of the bioreporter by the cellular population to the molecule capable of generating the detectable fluorescent signal correlates with rate of carbon dioxide production or rate of ethanol production by the cellular population. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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Specification