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Abstract
Flow cytometers can make thousands of single-cell measurements per second, but the limited ability to quantify uncertainty in individual measurements reduces their effectiveness for characterizing biomarker distributions, discriminating cell populations, and detecting rare events. To facilitate these analyses, we have developed a microfluidic flow cytometer that is capable of robust and repeated measurements with well-defined measurement uncertainty. We have invented and reduced to practice a device that can measure single objects in flow several times along a microchannel with integrated waveguides that deliver and collect emitted, transmitted, and scattered light. The invention builds on technology described NIST docket 18-017US1 (U.S. Patent Application serial number 15/967,966 “OPTICAL FLOW METER FOR DETERMINING A FLOW RATE OF A LIQUID,”), primarily as it describes method of exciting fluorophores in specific regions of microfluidic flow channel. This submission is also a primary component of another recently submitted DN-45 “Multiplex Amplitude Modulation Fluorometry” by the same inventors.
Features
Existing commercial systems do not enable repeat measurements, cannot account for uncertainty in measurement due to deformation or degradation, cannot easily separate groups of objects (e.g. doublets), and cannot easily discriminate similar objects. There is also measurement system capable of measuring dynamic behavior in high throughput (e.g. hundreds of cell per second).