Fluorescence Intensity Correction via Least Squares Estimation
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Solution Overview
Problem
Existing fluorescence correction methods, such as those using inverse matrices, can produce negative fluorescence intensity values and fail to accurately account for autofluorescence variations across sub-populations, leading to measurement errors and incorrect representation of population sizes in cytograms.
Innovation Solution
An information processing apparatus and method that express light intensity distributions from multi-dyed microparticles as a linear combination of reference spectra, modeled to follow a predetermined probability distribution, to estimate combination coefficients and correct fluorescence intensities based on fluorescence characteristics of single pigments, ensuring non-negative intensity values and improved accuracy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If inverse matrix method is used for fluorescence correction, then correction can be performed mathematically, but negative intensity values are produced leading to measurement errors
Solution Approach 1:
The patent changes the mathematical approach from inverse matrix method to least squares estimation. This parameter change in the correction method transforms the problem solution space, ensuring that intensity values remain non-negative while maintaining correction accuracy. The least squares method inherently provides non-negative solutions when proper constraints are applied, resolving the contradiction between mathematical correctness and physical validity of intensity values.
Solution Approach 2:
The patent replaces the traditional matrix inversion approach with a statistical estimation approach using least squares. This substitution of the mathematical mechanism allows for more robust handling of noise and ensures physically meaningful results. The least squares estimation with constraints provides a more reliable method that maintains both accuracy and the non-negativity requirement of intensity measurements.
2Ease of manufacture
If average autofluorescence intensity is used for correction, then calculation is simplified, but sub-population-specific variations are not accounted for causing errors
Solution Approach 1:
The patent segments the population into sub-populations based on fluorescence characteristics and applies correction calculations separately to each sub-population. This segmentation allows the system to account for sub-population-specific autofluorescence variations while maintaining computational feasibility. By dividing the complex correction problem into smaller, manageable sub-problems, the system achieves both accuracy and computational efficiency.
Solution Approach 2:
The patent applies local quality by tailoring the correction parameters to specific sub-populations rather than using a uniform approach. Each sub-population receives correction treatment customized to its specific autofluorescence characteristics. This localized approach ensures high precision for each sub-group while the overall system remains computationally manageable through structured processing.
3Measurement precision
If fluorescence correction is applied to multi-color measurement data, then measurement accuracy improves, but computational complexity increases
Solution Approach 1:
The patent performs preliminary actions by pre-calculating and storing correction parameters for different sub-populations and fluorescence combinations. This preliminary processing reduces the computational burden during actual measurement correction. By preparing correction data in advance, the system can quickly apply corrections during measurement without requiring complex real-time calculations, thus reducing operational complexity while maintaining high precision.
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 more accurate extraction of target spectrum components by preventing negative intensity values and accounting for autofluorescence variations, enhancing the precision of fluorescence intensity corrections and cytogram representations.
Implementation Method 1
an apparatus (for example, a flow cytometer) that irradiates a laser beam to microparticles marked by a fluorescent pigment and measures the intensity or a pattern of fluorescence coming from the excited fluorescent pigment
Implementation Method 2
a transparent wavelength band of an optical filter provided in each optical detector to restrict the light reception wavelength band is designed in accordance with the fluorescent wavelength of the fluorescence coming from the fluorescent pigment to be measured
Data Source
AI summary
Provided is an information processing apparatus including an estimation unit that expresses a light intensity distribution, which is obtained by irradiating light to a measurement object of a measurement target having a plurality of substances with mutually different responsive characteristics to the light on a surface and/or an inside of the measurement object, as a linear combination of light intensity distributions, which are obtained by irradiating the light to reference measurement objects, each of which has a single substance, models the light intensity distribution obtained from each of the reference measurement objects so as to follow a predetermined probability distribution, and estimates a combination coefficient of the linear combination from the light intensity distribution obtained from the measurement object of the measurement target.