Kill event optimization
Inventors
SAVAGE, Frederick • Ebersole, Matthew • McAda, Daniel • Appleyard, David • Xia, Zheng
Assignees
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Publication Number
US-11513114-B2
Publication Date
2022-11-29
Expiration Date
Abstract
A method of choosing which undesired cell to destroy in a multi-cell fluorescent event includes detecting fluorescence of cells, converting photons detected in the fluorescence into an analog voltage output signal, and identifying at least two discernable peaks associated with the cells. By looking solely at properties measured within the multi-cell fluorescent event, a decision of which cell to target for elimination can be made. Using this method with large population sizes can result in an effective sex skewed product. The sex skewed product can, for example, be formed from bull semen which is then later used to inseminate cows which results in an increased likelihood of giving birth to female cattle.
Core Innovation
The invention relates to a method of selecting which undesired cell to destroy in a multi-cell fluorescent event. The method detects a fluorescence of at least two cells physically spaced close enough that the at least two cells create one fluorescent event, and converts photons detected in the fluorescence of the at least two cells into an analog voltage output signal. The analog voltage output signal is analyzed by identifying a first peak and associating the first peak with a first cell, and identifying a second peak and associating the second peak with a second cell.
The method further measures an amplitude of the first peak and an amplitude of the second peak. Based on those measured amplitudes, the method defaults a kill laser to fire a kill beam at any cells having an amplitude less than an amplitude of another cell. The selection rule therefore uses peak amplitude comparison between first and second peaks to decide which undesired cell is destroyed within the multi-cell fluorescent event.
The disclosed approach also includes conditional refinements associated with cell-death selection logic, including a dead-cell threshold and an amplitude-difference comparison concept used to override the default firing rule. Additional signal processing refinements are described as filtering and digitizing the analog voltage output signal. The disclosed method is applied to form a sexed semen product, including in the context of bovine insemination described as bull semen.
Claims Coverage
The provided independent claim sets out a multi-cell fluorescent-event workflow with four main inventive features: fluorescence detection for closely spaced cells, analog voltage conversion and peak identification, peak amplitude measurement and association to cells, and default kill-laser selection based on amplitude comparison (with dependent claims introducing a dead-cell threshold override and additional analog signal conditioning, plus a sexed semen product outcome).
Selecting which undesired cell to destroy in a multi-cell fluorescent event by peak amplitude comparison
Defaulting a kill laser to fire a kill beam at any cells having an amplitude less than an amplitude of another cell after detecting a fluorescence of at least two cells creating one fluorescent event, converting photons into an analog voltage output signal, identifying a first peak and associating it with a first cell, identifying a second peak and associating it with a second cell, and measuring the amplitude of the first peak and the amplitude of the second peak.
Dead-cell threshold and amplitude-difference override logic
Defining a dead-cell threshold and using a difference between the amplitude of the first peak and the amplitude of the second peak compared against the dead-cell threshold to optionally override the default kill-laser firing rule.
Filtering and digitizing the analog voltage output signal
Including filtering and digitizing of the analog voltage output signal produced from converting photons detected in the fluorescence.
Forming a sexed semen product by executing the cell-destruction selection method
Forming a sexed semen product by executing the method of claims 11, 12, or 13.
Overall, the claim coverage centers on selecting which undesired cell to destroy within a single multi-cell fluorescent event by converting detected photons to an analog voltage signal, identifying first and second peaks associated with respective cells, measuring their amplitudes, and defaulting the kill laser to fire based on amplitude comparison, with dependent refinements including a dead-cell threshold override, filtering/digitizing of the analog voltage signal, and producing a sexed semen product.
Stated Advantages
Improved purity and yield in the reported performance tests.
More effective sex skew in a sexed semen product for bovine insemination.
Documented Applications
Producing a sexed semen product for bovine insemination, including use with bull semen and female/male cattle insemination as described in the partial content.
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