Multispectral Calibration via Mixed Dye Formulations
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Solution Overview
Problem
Multispectral analysis systems face challenges in accurately quantifying multiple fluorophores due to spectral cross-talk, which impairs the recovery of accurate quantitative information from assays.
Innovation Solution
The use of mixtures of calibration dyes in a single calibration plate well to calibrate multispectral imaging systems, reducing or eliminating cross-talk among spectral channels and enhancing spectral multiplexing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional calibration methods using pure dyes are used, then the calibration process is simple, but spectral cross-talk impairs recovery of accurate quantitative information
Solution Approach 1:
The patent combines multiple calibration dyes into single calibration plate wells, creating mixed dye formulations that simultaneously calibrate multiple spectral channels. This merging approach reduces the number of separate calibration steps needed while improving measurement accuracy by accounting for spectral cross-talk between adjacent fluorophores.
Solution Approach 2:
The patent performs preliminary calibration using mixed dye formulations that pre-account for spectral cross-talk effects. By incorporating interfering spectral contributors into the calibration mixtures beforehand, the system establishes correction factors that compensate for cross-talk during subsequent sample analysis, improving quantitative accuracy without adding complexity during actual measurement.
2Adaptability or versatility
If spectral channels are closely spaced to increase multiplexing capacity, then more fluorophores can be analyzed, but cross-talk increases and impairs accurate detection
Solution Approach 1:
The patent merges multiple calibration dyes representing different spectral contributors into single calibration wells. This allows the system to simultaneously characterize and correct for cross-talk between closely spaced spectral channels, enabling higher multiplexing capacity while maintaining the ability to distinguish between overlapping fluorophores through the mixed calibration formulations.
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 allows for the reduction of background contributions from interfering spectral contributors, leading to more accurate and precise analysis of fluorophores with overlapping emission spectra.
Implementation Method 1
a first fluorescent entity and a second fluorescent entity in the composition, wherein a fluorescence emission spectrum of the first fluorescent entity at least partially overlaps with a fluorescence emission spectrum of the second fluorescent entity
Data Source
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AI summary
Methods for calibrating a multispectral analysis system (1000) include calibrating the system to detect fluorescence emission from a first fluorescent entity in a biological sample (900) that includes the first fluorescent entity and a second fluorescent entity using a calibration sample, where the calibration sample features a first concentration of the first fluorescent entity and a second concentration of the second fluorescent entity, and where the first concentration is larger than the second concentration.