Fluorescence Lifetime Analysis for Particle Autofluorescence Discrimination
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Solution Overview
Problem
Current multiplexed particle and cell analysis techniques face challenges with spectral overlap and autofluorescence, limiting the number of concurrent spectral bands and analytes that can be detected, and are often destructive or not suited for sorting.
Innovation Solution
The use of fluorescence lifetime as an independent parameter, combined with spectral fluorescence labeling, to reduce spectral crosstalk and autofluorescence interference, enabling more multiplexed combinations and non-destructive analysis and sorting.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If spectral fluorescence labeling is used for multiplexed analysis, then the number of detectable analytes increases, but spectral overlap and autofluorescence interference increase
Solution Approach 1:
The patent transitions from single-parameter spectral detection to multi-parameter analysis by incorporating fluorescence lifetime as an additional dimension. This allows discrimination between fluorophores based on both wavelength and temporal characteristics, enabling resolution of spectral overlaps and autofluorescence interference that cannot be resolved by wavelength alone.
Solution Approach 2:
The patent segments the fluorescence signal into distinct temporal components using lifetime gating. By dividing the detection window into early and late time segments, the system separates autofluorescence (short lifetime) from exogenous fluorophores (long lifetime), enabling selective detection of specific analytes while rejecting interfering signals.
2Adaptability or versatility
If multiple spectral bands are used simultaneously, then multiplexing capability increases, but device complexity increases
Solution Approach 1:
The patent implements a single fluorescence detector that performs multiple functions: spectral discrimination, lifetime measurement, and multiplexed analyte detection. This universal detector replaces the need for multiple specialized detection systems, achieving high multiplexing capability without proportionally increasing device complexity.
Solution Approach 2:
The patent replaces complex mechanical spectral separation systems with electronic signal processing. Instead of using multiple physical filters and detectors, the system uses digital signal analysis to distinguish between different fluorophores based on their temporal decay characteristics, simplifying the overall device architecture while maintaining high multiplexing capability.
3Measurement precision
If destructive analysis methods are used, then detection sensitivity increases, but sorting capability is lost
Solution Approach 1:
The patent employs a non-destructive fluorescence lifetime measurement technique that allows the same sample to be analyzed multiple times without degradation. The measurement process itself serves as the detection mechanism, preserving the sample integrity and enabling subsequent sorting operations without sacrificing detection sensitivity.
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 increases the number of distinguishable bead types and analytes, reduces costs, and allows for highly multiplexed, non-destructive analysis and sorting with reduced spectral overlap and autofluorescence interference.
Implementation Method 1
The use of fluorescence lifetime as an independent parameter, combined with spectral fluorescence labeling
Implementation Method 2
a detector comprising a number of spectral detection channels sensitive to distinct wavelength sections of the electromagnetic spectrum, such that the channels are configured to detect optical signals resulting from interactions between the beam and the sample and convert the optical signals into respective electrical signals
Data Source
AI summary
Described herein are apparatuses and methods for analyzing an optical signal decay. In some embodiments, an apparatus includes: a source of a beam of pulsed optical energy; a sample holder configured to expose a sample to the beam; a detector comprising a number of spectral detection channels configured to convert the optical signals into respective electrical signals; and a signal processing module configured to perform a method. In some embodiments, the method includes: receiving the electrical signals from the detector; mathematically combining individual decay curves in the electrical signals into a supercurve, the supercurve comprising a number of components, each component having a time constant and a relative contribution to the supercurve; and quantifying a relative contribution of each component to the supercurve.