Flow Cytometer Fluorochrome Detection Using Spillover Coefficients
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Solution Overview
Problem
The complexity and cost of multi-laser, multi-detector flow cytometers make them inaccessible for multi-color and multi-parameter analysis, as they require numerous detectors for each parameter measured, increasing size and operational complexity.
Innovation Solution
A method and system that utilize distinct spillover coefficients of fluorochromes to detect multiple fluorochromes with fewer detectors by characterizing and sorting particles based on spectral spillover coefficients, allowing for analysis with two detectors instead of one for each fluorochrome.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If multiple lasers and detectors are added to measure more fluorescence signals simultaneously, then the multi-parameter analysis capability is improved, but the size and complexity of the flow cytometer increases significantly
Solution Approach 1:
The patent makes each detector universal by enabling it to detect multiple fluorochromes simultaneously through spectral unmixing. Instead of requiring one detector per fluorochrome, a single detector can collect signals from multiple fluorochromes with distinct spillover coefficients, and computational algorithms separate the individual signals. This multi-functionality reduces the total number of detectors needed while maintaining the ability to analyze multiple parameters.
Solution Approach 2:
The patent changes the detection parameter from wavelength-specific detection to spillover coefficient-based detection. By characterizing fluorochromes based on their unique spillover coefficients rather than their emission wavelengths alone, the system can distinguish between multiple fluorochromes using fewer detectors. The spectral unmixing algorithms use these parameter changes to mathematically separate overlapping fluorescence signals.
2Adaptability or versatility
If multiple lasers and detectors are added to measure more fluorescence signals simultaneously, then the multi-color analysis capability is improved, but the cost and difficulty of setting up, operating, and maintaining the system increases
Solution Approach 1:
The patent makes each detector universal by enabling it to detect multiple fluorochromes simultaneously through spectral unmixing. Instead of requiring one detector per fluorochrome, a single detector can collect signals from multiple fluorochromes with distinct spillover coefficients, and computational algorithms separate the individual signals. This multi-functionality reduces the total number of detectors needed while maintaining the ability to analyze multiple parameters.
3Device complexity
If the number of detectors is reduced to decrease system complexity, then the accessibility and ease of use is improved, but the ability to detect multiple fluorochromes is compromised
Solution Approach 1:
The patent introduces computational algorithms as an intermediary between the detector and the final measurement result. The detector collects mixed fluorescence signals from multiple fluorochromes, and the spectral unmixing algorithms act as intermediaries to mathematically separate and identify individual fluorochrome signals based on their unique spillover coefficient patterns. This intermediary processing layer enables accurate multi-fluorochrome detection with fewer physical detectors.
Solution Approach 2:
The patent replaces the mechanical approach of using separate detectors for each fluorochrome with a computational approach. Instead of physically separating detection paths with multiple detectors, the system uses mathematical algorithms to separate fluorochrome signals computationally. This substitution of mechanical complexity with computational processing maintains detection accuracy while reducing the number of physical components.
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 reduces the number of detectors needed, making flow cytometry more accessible and user-friendly while maintaining high-quality data capture across a wide range of fluorescence intensities, ensuring consistent and reproducible results.
Implementation Method 1
the parameters that are analyzed include light scatter and fluorescence signals, generated by interaction of the particles with light sources in the flow cytometer
Implementation Method 2
detecting the first and second fluorochromes of the particles with a first detector and a second detector
Data Source
AI summary
A method for detecting fluorochromes in a flow cytometer, including: receiving a sample including particles each tagged with at least one of a first fluorochrome and a second fluorochrome, in which the first and second fluorochromes having distinct spillover coefficients; detecting the particles, including detecting the first and second fluorochromes with a first detector and a second detector; forming a data set for detected particles based on the detection of the first and second fluorochromes; characterizing a detected spillover coefficient for each detected fluorochrome from the data set; and sorting the detected particles into predicted fluorochrome populations based on the detected spillover coefficients. A system for detecting fluorochromes in a flow cytometer, including a flow cell, a fixed gain detection system, and a processor that generates a detected spillover coefficient for each detected particle and sorts the detected particle into predicted fluorochrome populations based on the detected spillover coefficient.