Metabolic rate indicator for cellular populations
View Patent ↗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.
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.
2. The method of claim 1 , wherein the cellular population is selected from bacteria, archae, protists, microscopic animals, fungi, yeast, microscopic plants, animal cells, and any combination of the foregoing.
3. The method of claim 1 , wherein the sample of the cellular population is obtained via a manually operated sampler.
4. The method of claim 1 , wherein the sample containing the cellular population undergoes separation and resuspension.
5. The method of claim 1 , wherein the method is carried out via a vessel comprising the cellular population, a sample collection port, a sample line, and a joint between the sample collection port and the sample line, wherein the joint includes a means to create a fluid communication channel between the sample collection port and the sample line so as to allow a sample of the cellular population to be drawn into the sample line, wherein the sample collection port is in communication with the vessel, wherein the sample line is in communication with the vessel via a valve.
6. The method of claim 5 , wherein the metabolic signal is detected via a fluorometer located at least partially inside the vessel.
7. The method of claim 1 , further comprising combining additive with the sample containing the cellular population, the bioreporter, and/or combination thereof, wherein the additive causes the parameter to change when combined with the sample, the bioreporter, and/or combination thereof, thereby constituting the initial point of reaction.
8. The method of claim 7 , wherein the additive is selected from a buffering agent, a pH modifying agent, a catalyst, a coenzyme, a mineral, a co-substrate, and combinations thereof.
9. The method of claim 7 , wherein the additive is a buffering agent that buffers the sample to a pH of approximately 7.6.
10. The method of claim 1 , wherein the sample of the cellular population is obtained via an automatically operated sampler.
11. The method of claim 1 , wherein the sample of the cellular population is obtained via a sidestream apparatus.
12. The method of claim 1 , wherein the sample containing the cellular population is diluted.