Cell differentiation based on multi-directional light from a microfluidic chip
Inventors
Appleyard, David • McAda, Daniel • Xia, Zheng • Schiller, James Maxwell • SAVAGE, Frederick • Rupel, II, John Walker • Miller, Timothy • Fisher, Alec
Assignees
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Publication Number
US-12350665-B2
Publication Date
2025-07-08
Expiration Date
Abstract
Disclosed is an approach to differentiating between different particle types in samples flowing through microfluidics chips. A sample may have an initial proportion of a first cell type to a second cell type. An illuminating light source may emit a coherent light at the sample, and light leaving the chip in a first direction may be detected using a first light detector, and light leaving the chip in a second direction (e.g., orthogonal to the first direction) may be detected using a second light detector. The detected light may be fluorescence. An orientational feature of a plurality of cells in the sample may be determined based on the light detected by the detectors. Based on the orientational features and the detected light, a biasing operation may be performed for each cell in the sample to obtain a modified proportion of cell types in the sample.
Core Innovation
The disclosure relates to a method using a microfluidic chip having a sample channel to process a sample comprising a particle population having a proportion of a first particle type to a second particle type. The method emits illuminating light from a light source along a first axis such that the illuminating light coincides with particles as the sample passes through the channel, and detects fluorescence emitted along a first axis and a second axis using one or two light detectors.
The first light and the second light are fluorescence that results from incidence of the illuminating light on particles in the sample and that travels in multiple different directions from fluorescing particles. Based on the detected first light and the detected second light, the method performs a biasing operation that modifies the proportion of the first particle type to the second particle type, and the fluorescence data are used to generate an intensity map of peak fluorescence associated with each particle type.
The method further defines particle selection gates by locating population centers for the first particle type and the second particle type, calculating relative intensity along a line passing through a population center, calculating a slope of a line joining the population centers, and locating saddle points along lines joining corresponding portions of the particle types. The method also generates a gate by bounding a population using the saddle points and additional bounding points, or by processing a pixel map with kernel-based computations to derive histograms, trough points, and a polynomial function for bounding a population.
Claims Coverage
The document provides three independent methods. Each independent claim specifies a microfluidic, fluorescence-based detection along a first and a second axis, followed by a biasing operation to modify the proportion between two particle types, and includes additional data-processing steps to define how populations are bounded.
Microfluidic axis-based fluorescence detection and biasing of proportions
Providing a sample to a microfluidic chip with a sample channel, illuminating particles along a first axis, detecting fluorescence emitted along a first axis and a second axis using one or two light detectors, and performing a biasing operation based on the detected fluorescence along the first and second axes to modify the proportion of a first particle type to a second particle type, wherein the fluorescence travels in multiple different directions from fluorescing particles.
Intensity map with population centers, lines, saddle points, and gate bounding
Generating an intensity map of peak fluorescence associated with each particle type, detecting a first population center and a second population center, calculating relative intensity along a first line passing through the first population center, calculating a slope of a second line joining the population centers, locating first and second saddle points along third and fourth lines joining corresponding populations, and generating a gate by bounding one of the populations based on the saddle points and additional bounding points.
Pixel map kernel processing to derive histograms, trough points, and polynomial bounding
Calculating a kernel within a pixel map and adding instances of the kernel based on light emitted from the sample along the first and second axes, deriving histograms from the pixel map, generating trough points from the histograms, generating a polynomial function based on the trough points, and bounding a population based on the polynomial function and additional bounding points.
Across the independent claims, the core inventive coverage combines microfluidic axis-based fluorescence detection with multi-directional fluorescence and a biasing operation that modifies the proportion of two particle types, together with population-bounding logic using intensity-map population-center, line, and saddle-point gate generation or pixel-map kernel, histogram, trough-point, and polynomial processing.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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