Microfluidic system and method with focused energy apparatus
Inventors
Appleyard, David • BETTHAUSER, Jeff • FAUST, Marjorie • Larsen, John • SHAO, Guocheng • Xia, Zheng • Zhou, Yu
Assignees
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Publication Number
US-11639888-B2
Publication Date
2023-05-02
Expiration Date
Abstract
A method of inseminating an animal including flowing a stream of a population of sperm cells through a channel, differentiating the sperm cells into two subpopulations of X-chromosome containing sperm cells and Y-chromosome containing sperm cells, selecting a desired subpopulation, ablating an undesired subpopulation, and collecting both the subpopulations of sperm cells including the desired subpopulation and the ablated undesired subpopulation together, wherein the collected population of sperm cells is used to fertilize an egg.
Core Innovation
The invention relates to a flow cytometry apparatus configured to produce a sexed composition of non-human animal sperm by identifying a subpopulation of sperm from a population of sperm in a sample fluid. The apparatus includes an interrogation light source, a detector, and computer readable media with instructions that define and store a gating strategy. The gating strategy differentiates the subpopulation based on physical characteristics of sperm that are identifiable based in part on interrogation by the interrogation light source as detected by the detector.
The apparatus measures one or more physical characteristics for the sperm in the population based on a set of signals detected by the detector. The gating strategy is updated based on the measured one or more physical characteristics, and the updated gating strategy is stored. Using the measured one or more physical characteristics and the updated gating strategy, the apparatus identifies sperm in the subpopulation of sperm.
The document also describes microfluidic flow-through and focused-energy actions used with sperm subpopulation identification to ablate an undesired X- or Y-sperm subpopulation, thereby enabling sexed sperm composition outcomes and collection options for desired and ablated sperm.
Claims Coverage
The document includes two independent claims covering a flow cytometry apparatus that performs gating strategy definition, updating, and subpopulation identification to produce a sexed sperm composition, and non-transitory computer readable media that causes a flow cytometry instrument to implement a gating strategy workflow for producing a sexed sperm composition.
Gating strategy definition, updating, and sperm subpopulation identification
A flow cytometry apparatus configured to produce a sexed composition of non-human animal sperm by identifying a subpopulation of sperm from a population of sperm in a sample fluid, wherein the apparatus defines and stores a gating strategy for differentiating the subpopulation based on one or more physical characteristics of sperm in the population identifiable based in part on interrogation by the interrogation light source as detected by the detector; measures the one or more physical characteristics based on a set of signals detected by the detector; updates the gating strategy based on the one or more physical characteristics measured; stores the updated gating strategy; and identifies sperm in the subpopulation based on the measured one or more physical characteristics and the updated gating strategy.
Non-transitory computer readable media for implementing a gating strategy workflow
Non-transitory computer readable media for use with a flow cytometry instrument, comprising instructions that cause the flow cytometry instrument to define and implement a gating strategy for producing a sexed composition of sperm comprising non-human animal sperm by identifying a subpopulation of sperm from a population of sperm in a sample fluid, wherein the instructions measure one or more physical or genetic characteristics present in the sexed composition of sperm based on an output of a detector; set gating parameters for differentiating the subpopulation based on the one or more physical or genetic characteristics and wherein the gating parameters comprise marking one or more desired physical or genetic characteristics of the subpopulation; store the defined gating strategy; measure the one or more physical or genetic characteristics for the sperm based on the defined gating strategy; update the defined gating strategy based on the measured one or more physical or genetic characteristics to generate an updated gating strategy; store the updated gating strategy; and identify sperm in the subpopulation based on the measured one or more physical or genetic characteristics and the updated gating strategy.
Across the independent claims, the inventive coverage centers on defining a gating strategy to differentiate a sperm subpopulation based on measured detector-interpretable characteristics, updating and storing that gating strategy based on measured characteristics, and identifying sperm in the gated subpopulation to produce a sexed composition of non-human animal sperm. A second independent claim additionally covers non-transitory computer readable media that implements this gating workflow on a flow cytometry instrument, including measurement, marking desired characteristics, gating parameter setting, updating, and identification using the updated gating strategy.
Stated Advantages
Produces a sexed composition of sperm comprising non-human animal sperm by identifying a sperm subpopulation in a sample fluid using a gating strategy and an updated gating strategy.
Enables gating strategy adaptation by updating the gating strategy based on measured one or more physical or genetic characteristics.
Supports identification of sperm in the desired subpopulation by using measured characteristics together with the updated gating strategy.
Documented Applications
Sexed sperm for non-human animal sperm, including cattle (heifer/cow) conception-rate comparisons for sexed insemination using the described approach versus conventional unsexed and competitor physically separated sexing technologies [procedural detail omitted for safety].
A broader discrimination context for biomedical particle discrimination using focused-energy actions (kill/disable/alter/destroy, photobleach, photopolymerize) in combination with identification of subpopulations based on interrogation.
Application to other cells/biomedical particles is described as broadly applicable to cellular applications including stems cells, labeled cells, blood cells, viruses/bacteria/organelles [procedural detail omitted for safety].
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