Fluorescence Intensity Correction Using Pre-Calculated Matrices
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Solution Overview
Problem
Current fluorescence intensity correction methods require analyzing singly labeled microparticles for every sample analysis, which is time-consuming and prone to measurement errors, especially when using multiple fluorescent pigments, as they often involve manual corrections and storage of reference data that does not fully eliminate measurement errors.
Innovation Solution
A method that separates the fluorescence spectrum of multiply labeled microparticles into small spectrum groups using independent component analysis or principal component analysis, compares these groups with pre-obtained fluorescence wavelength distributions to specify the spectrum of each fluorescent pigment, and calculates the intensity using a correction matrix, allowing for accurate fluorescence intensity calculation without needing to analyze singly labeled microparticles for every sample.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorescence correction is performed by analyzing singly labeled microparticles for every sample analysis, then measurement precision is improved, but loss of time increases due to repeated preparation and analysis
Solution Approach 1:
The patent creates a correction matrix in advance by analyzing singly labeled microparticles once, storing the fluorescence intensity relationships between multiple pigments. This preliminary correction matrix is then reused for correcting multiply labeled sample measurements without requiring repeated singly labeled analyses, thus resolving the contradiction between measurement precision and time loss.
2Loss of information
If the number of fluorescent pigments is increased for more detailed analysis, then information completeness is improved, but device complexity increases due to more correction calculations
Solution Approach 1:
The patent pre-calculates and stores a correction matrix that encapsulates the fluorescence intensity relationships among multiple pigments. This allows the system to handle an increased number of fluorescent pigments for more detailed sample analysis without proportionally increasing calculation complexity during actual sample measurement, as the correction matrix reusable for all subsequent measurements.
Solution Approach 2:
The patent creates a mathematical model (correction matrix) that represents the fluorescence relationships among multiple pigments. This model serves as a simplified copy or representation of the complex fluorescence interactions, allowing accurate correction of multiply labeled samples without directly computing complex relationships each time, thus managing device complexity while supporting multiple pigments.
3Ease of operation
If reference fluorescence wavelength distribution is stored in the device in advance, then ease of operation is improved, but measurement precision deteriorates because measurement errors for every analysis cannot be removed
Solution Approach 1:
The patent performs preliminary analysis of singly labeled microparticles to create a correction matrix that captures actual fluorescence intensity relationships. While this requires initial setup, it enables accurate correction for every subsequent sample analysis by accounting for specific measurement conditions, thus improving measurement precision while maintaining ease of operation through automated correction.
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 simplifies and accurately calculates fluorescence intensity from each fluorescent pigment, reducing measurement errors and preparation time, enabling more detailed analysis with a greater number of fluorescent pigments without the need for extensive pre-analysis preparation.
Implementation Method 1
measuring the properties of microparticles by labeling microparticles such as cells using fluorescent pigments and measuring the intensity and the pattern of the fluorescence emitted from the fluorescent pigments that are excited by being irradiated by laser light
Data Source
AI summary
A fluorescence intensity correction method including: receiving fluorescence emitted from fluorescent pigments, collecting detected values from each light detector, and obtaining a fluorescence spectrum of each fluorescent pigment as one spectrum group; separating the obtained spectrum group into a plurality of small spectrum groups; comparing the separated small spectrum groups with a fluorescence wavelength distribution of each fluorescent pigment obtained in advance and specifying the small spectrum groups as the fluorescence spectrum of any fluorescent pigment; comparing a differential spectrum between small spectrum groups that are not specified and one or more specified small spectrum groups with the fluorescence wavelength distribution of unspecified fluorescent pigments obtained in advance, and specifying the differential spectrum as the fluorescence spectrum of any fluorescent pigment; and calculating the intensity of fluorescence emitted from each fluorescent pigment.