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US 10,502,678 B2Granted
Systems and methods for panel design in flow cytometry
AssigneeBeckman Coulter, Inc.
InventorMichael Kapinsky
Life of this patent
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Abstract
Embodiments of the present invention encompass systems and methods for determining detection limits for various antibody-dye conjugates for flow cytometry. Exemplary techniques involve a linear superpositioning approach of spillover-induced enlargements of normally distributed measurement errors.
Claims20
A method of designing a probe panel for differentiating populations of particles in a sample using a flow cytometer, the method comprising:
- determining a distortion factor that quantifies spillover effect caused by emission of a first label, intended to be measured in a first channel, into a second channel;
- determining a maximum expected signal of a first probe-label combination including the first label and a first probe, wherein the first probe binds to a first antigen associated with a population of particles of interest in the sample, and wherein the maximum expected signal is determined based on a brightness of the first label and a density of the first antigen on or inside the particles of interest in the sample;
- calculating in a detection limit of a second antigen detected in the second channel, wherein calculating the detection limit comprises multiplying the distortion factor by the maximum expected signal of the first probe-label combination, and wherein the detection limit is a threshold of signal intensity dividing particles considered to be positive for the second antigen from particles considered to be negative for the second antigen;
- selecting a probe-label combination to include in the probe panel based on the calculated detection limit;
- and
- differentiating the populations of particles in the sample using the probe panel in the flow cytometer.
3 more elements
A method of designing a probe panel for differentiating populations of particles in a sample using a flow cytometer, the method comprising:
- determining a distortion factor that quantifies spillover effect caused by emission of a first label, intended to be measured in a first channel, into a second channel;
- determining a maximum expected signal of a first probe-label combination including the first label and a first probe, wherein the first probe binds to a first antigen associated with a population of particles of interest in the sample, and wherein the maximum expected signal is a function of a brightness of the first label and an expression pattern of the first antigen;
- calculating a detection limit of a second antigen detected in the second channel, wherein calculating the detection limit comprises multiplying the distortion factor by the maximum expected signal of the first probe-label combination, and wherein the detection limit is a threshold of signal intensity dividing particles considered to be positive for the second antigen from particles considered to be negative for the second antigen;
- selecting a probe-label combination to include in the probe panel based on the calculated detection limit;
- and
- differentiating the populations of particles in the sample using the probe panel in the flow cytometer.
3 more elements
A method of designing a probe panel for differentiating populations of particles in a sample using a flow cytometer, the method comprising:
- determining a distortion factor that quantifies spillover effect caused by emission of a first label, intended to be measured in a first channel, into a second channel;
- determining a maximum expected signal of a first probe-label combination including the first label and a first probe, wherein the first probe binds to a first antigen associated with a population of particles of interest in the sample, and wherein the maximum expected signal is a function of the brightness of the first label and an expression pattern of the first antigen;
- calculating a detection limit of a second antigen detected in the second channel using a second probe-label combination including a second label and a second probe corresponding to the second antigen, wherein calculating the detection limit comprises multiplying the distortion factor by the maximum expected signal of the first probe-label combination, and wherein the detection limit is represented as a slope of a graph of emission measured in the second channel as a function of emission measured in the first channel;
- selecting a probe-label combination to include in the probe panel based on the calculated detection limit;
- and
- differentiating the populations of particles in the sample using the probe panel in the flow cytometer.
3 more elements
Dependencies are inferred from the claim text.