Multi-photon counting flow cytometer detection sensitivity
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Solution Overview
Problem
Current methods for characterizing analytes in biological fluids, such as flow cytometry, face limitations in accurately detecting and quantifying light signals from samples, particularly in terms of intensity and sensitivity, which can impact disease diagnosis and treatment protocols.
Innovation Solution
The implementation of multi-photon counting techniques, which involve irradiating a sample in a flow stream with a light source and detecting photons by integrating photo-electron charge over specific time intervals, allowing for enhanced detection and quantitation of light signals through digital and analog output signals.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If single photon counting is used for light detection, then the system is simple to operate, but the detection sensitivity and quantitation range are limited
Solution Approach 1:
The patent divides the detection process into discrete time intervals (e.g., 100 μs intervals) and segments the photon counting into multiple channels (single photon counter and multi-photon counter). This segmentation allows the system to handle different light intensity ranges separately, improving overall measurement precision while managing complexity through modular processing.
Solution Approach 2:
The patent introduces a temporal dimension by implementing time-gated detection with multiple time intervals. Instead of measuring all photons simultaneously, the system counts photons in sequential time windows, effectively adding a time dimension to the detection process. This broadens the quantitation range by 100-fold while maintaining system feasibility.
2Measurement precision
If longer time intervals are used for photon counting, then more photons are detected improving sensitivity, but the time resolution between sequential measurements decreases
Solution Approach 1:
The patent segments the total measurement time into multiple discrete time intervals (e.g., ten 100 μs intervals). Each interval contributes to the total photon count, improving sensitivity through accumulated photons, while the segmentation maintains time resolution by clearly defining start and end points for each measurement window.
Solution Approach 2:
The patent implements continuous photon counting across multiple time intervals without gaps between measurements. The detector continuously integrates photoelectron charge throughout the sequence of time intervals, ensuring no useful signal is lost while accumulating sufficient photons for high sensitivity detection.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach significantly broadens the range of light intensity detection and quantitation, improving the accuracy of characterizing sample components, including extracellular vesicles, by up to 100-fold compared to single photon counting, facilitating more precise medical diagnoses and assessments.
Implementation Method 1
detecting light from the sample in the flow stream and counting photons of the detected light by integrating photo-electron charge over a time interval
Data Source
AI summary
Aspects of the present disclosure include methods and systems for detecting light from a sample in a flow stream by multi-photon counting. Methods according to certain embodiments include irradiating a sample in a flow stream with a light source and detecting light from the sample in the flow stream and counting photons of the detected light by integrating photo-electron charge over a time interval. Methods also include irradiating a sample in a flow stream with a light source, detecting light from the sample in the flow stream and outputting a digital output signal and an analog output signal produced by the detected light. Systems for detecting light from a sample in a flow stream with a detector and counting photons by integrating photo-electron charge over a time interval are also described. Kits having a detector, a photon counter and a flow cell configured to propagate a sample in flow stream are also provided.