Fluorescence Intensity Calculation Using Overdetermined Spectral Fitting
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Solution Overview
Problem
Existing fluorescence intensity correction methods are limited by the need for equal numbers of photodetectors and fluorescent dyes, leading to inefficiencies in data utilization and inaccurate intensity calculations when using more photodetectors than dyes in multicolor measurements.
Innovation Solution
A method that approximates measured spectra using a linear sum of single-dyeing spectra, employing a least-squares approach to calculate fluorescence intensities, and optionally using singular value decomposition to handle invalid data points, allowing for precise intensity calculations regardless of the number of fluorescent dyes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the number of photodetectors is increased to improve measurement precision in multicolor analysis, then data precision improves, but device complexity and data processing difficulty increase
Solution Approach 1:
The patent changes the mathematical approach from traditional matrix inversion to least-squares approximation with singular value decomposition, allowing the system to handle cases where the number of photodetectors exceeds the number of fluorescent dyes. This parameter change in the calculation method enables effective utilization of all photodetector data without requiring equal numbers of photodetectors and dyes.
2Speed
If fluorescence correction is performed using traditional matrix inversion methods, then intensity calculation speed improves, but measurement precision deteriorates when photodetector count exceeds dye count
Solution Approach 1:
The patent introduces a dynamic calculation approach using least-squares approximation that adapts to different configurations of photodetectors and fluorescent dyes. The method dynamically adjusts to handle cases where photodetector count exceeds dye count, maintaining both speed and precision through iterative optimization rather than fixed matrix inversion.
3Productivity
If all photodetector data is utilized to improve productivity, then data utilization efficiency improves, but calculation complexity increases
Solution Approach 1:
The patent extracts the essential fluorescence intensity information from all photodetector data through least-squares approximation, separating the useful signal from the complexity of having more photodetectors than dyes. By using singular value decomposition, the method extracts only the necessary components while discarding redundant information, achieving high data utilization without proportional increase in processing complexity.
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
Enables effective utilization of all photodetector data to precisely calculate fluorescence intensities from multiple dyes, enhancing data precision and separation accuracy in multicolor measurements.
Implementation Method 1
measuring an intensity or a pattern of a fluorescence generated from the fluorescent dye excited by radiating a laser beam to the microparticle
Data Source
AI summary
A fluorescence intensity calculating apparatus, includes a measuring section configured to receive fluorescences generated from plural fluorescent dyes excited by radiating a light to a microparticle multiply-labeled with the plural fluorescent dyes having fluorescence wavelength bands overlapping one another by photodetectors which correspond to different received light wavelength bands, respectively, and whose number is larger than the number of fluorescent dyes, and obtain measured spectra by collecting detected values from the photodetectors, and a calculating section configured to approximate the measured spectra based on a linear sum of single-dyeing spectra obtained from the microparticle individually labeled with the fluorescent dyes, thereby calculating intensities of the fluorescences generated from the fluorescent dyes, respectively.