Methods for sorting particles
First Claim
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1. A method of sorting particles in a flow cytometer comprising the steps of:
- a. establishing a mathematical model based upon extracted features of waveform pulses produced by stained particles when run through the flow cytometer, wherein the particles flowing through the flow cytometer belong to at least a first population or a second population, wherein the mathematical model represents a statistical distribution of extracted features of the waveform pulses;
b. analyzing the waveform pulses produced by stained particles flowing through the flow cytometer to determine a probability each particle belongs to the first population and the second population based upon the mathematical model;
c. assigning each stained particle a value based upon the probabilities associated with belonging to the first population or the second population;
d. establishing a decision boundary;
e. modifying the value assigned to each stained particle relative to the decision boundary based upon a purity or quantity of the first population or of the second population; and
f. separating the stained particles based upon their modified relationship relative to the decision boundary.
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Abstract
A method of sorting stained particles in a flow cytometer by generating a mathematical model based upon extracted features of waveform pulses produced in response to the stained particles. Waveform pulses can then be analyzed to determine the probability the stained particles belong in either a first or a second population and a decision boundary can be established. Sorting decisions can be made based upon the extracted features of the waveform pulses relative to the decision boundary in combination with other desirability factors for particles or droplets containing the particles.
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Citations
30 Claims
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1. A method of sorting particles in a flow cytometer comprising the steps of:
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a. establishing a mathematical model based upon extracted features of waveform pulses produced by stained particles when run through the flow cytometer, wherein the particles flowing through the flow cytometer belong to at least a first population or a second population, wherein the mathematical model represents a statistical distribution of extracted features of the waveform pulses; b. analyzing the waveform pulses produced by stained particles flowing through the flow cytometer to determine a probability each particle belongs to the first population and the second population based upon the mathematical model; c. assigning each stained particle a value based upon the probabilities associated with belonging to the first population or the second population; d. establishing a decision boundary; e. modifying the value assigned to each stained particle relative to the decision boundary based upon a purity or quantity of the first population or of the second population; and f. separating the stained particles based upon their modified relationship relative to the decision boundary. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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13. A method of sorting particles in a flow cytometer comprising the steps of:
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a. establishing a mathematical model based upon extracted features of waveform pulses produced by stained particles when the flow cytometer, wherein the particles flowing through the flow cytometer belong to at least a first population or a second population, wherein the mathematical model represents a statistical distribution of extracted features of the waveform pulses; b. analyzing the waveform pulses produced by stained particles flowing through the flow cytometer to determine a probability each particle belongs to the first population and the second population based upon the mathematical model; c. assigning each stained particle a value based upon the probabilities associated with belonging to the first population or the second population; d. establishing a decision boundary; e. modifying the value assigned to each stained particle relative to the decision boundary, wherein the modification of the value assigned to each particle is related to a cost associated with misidentifying a particle as belonging to the first population or the second population; and f. separating the stained particles based upon their modified relationship relative to the decision boundary. - View Dependent Claims (14)
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15. A method of sorting particles in a flow cytometer comprising the steps of:
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a. establishing a mathematical model based upon extracted features of waveform pulses produced by stained particles when run through the flow cytometer, wherein the particles flowing through the flow cytometer belong to at least a first population or a second population wherein the mathematical model represents a statistical distribution of extracted features of the waveform pulses; b. analyzing the waveform pulses produced by stained particles flowing through the flow cytometer to determine a probability each particle belongs to the first population and the second population based on the mathematical model; c. entraining the stained particles in droplets and determining the desirability of a droplet containing one or more of the stained particles; d. establishing a decision boundary relating to at least one extracted feature of the waveform pulse for a stained particle; and e. collecting droplets based upon the relationship of the one or more stained particles within each droplet relative to the decision boundary and based upon the determined desirability of the droplet. - View Dependent Claims (16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
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Specification