Fluorescent Particle Quantification via Luminance Thresholding
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Solution Overview
Problem
Conventional methods for quantitative determination of biological substances using fluorescent particles are inaccurate due to clustering of particles and background noise, leading to errors in calculating average luminance per particle.
Innovation Solution
A method involving inputting a fluorescent image, extracting a predetermined region, calculating integrated luminance, and counting fluorescent particles using a correlation between the number of particles and their luminance, with optional steps including bright spot extraction and scanning electron microscope counting, to accurately determine the number of specific biological substances.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the average luminance per phosphor particle is calculated from the fluorescent image by analyzing peaks in the luminance distribution, then the quantitative determination can be performed, but the calculation becomes inaccurate when phosphor particles form clusters and background noises are present
Solution Approach 1:
The patent extracts only the fluorescent signal components corresponding to phosphor particles by thresholding the luminance distribution, separating the useful signal from harmful background noises and clustered particle effects. This extraction enables accurate measurement of individual particle luminance despite the presence of clusters and noise in the original image.
Solution Approach 2:
The patent applies local analysis by calculating the luminance distribution characteristics specifically for regions containing phosphor particles, rather than using global average luminance. This local approach allows accurate determination of particle luminance even when particles are clustered, as the analysis focuses on the specific local characteristics of fluorescent signals.
2Measurement precision
If the number of fluorescent particles is determined by analyzing the fluorescent image, then quantitative determination of biological substance can be achieved, but background noises and clustering cause errors in the determination
Solution Approach 1:
The patent extracts fluorescent particle information by thresholding the luminance distribution to identify and separate true fluorescent signals from background noises. This extraction enables accurate counting of fluorescent particles even when they are clustered or when background noise is present, as the thresholding process selectively identifies only the fluorescent components.
Solution Approach 2:
The patent introduces the luminance distribution characteristics as an intermediary parameter to bridge the gap between raw fluorescent image data and accurate particle counting. By analyzing the luminance distribution properties, the system can accurately determine particle number despite the interfering effects of clustering and background noise.
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 method enables precise quantification of biological substances by reducing errors from particle clustering and background noise, providing accurate results for pathological diagnosis support systems.
Implementation Method 1
fluorescent particles each encapsulating a fluorescent substance and binding to a biological substance recognizing site
Data Source
AI summary
A quantitative determination method of a biological substance in a sample stained with a staining reagent comprising fluorescent particles each encapsulating a fluorescent substance and binding to a biological substance recognizing site. The method comprises inputting a fluorescent image obtained by photographing the sample, extracting a predetermined region from the fluorescent image to calculate an integrated luminance of the predetermined region, and counting the number of fluorescent particles contained in the predetermined region from the integrated luminance and the average luminance per fluorescent particle. The average luminance per fluorescent particle is calculated from a correlation between the number of fluorescent particles counted from an image of the fluorescent particles visualized and the luminance derived from fluorescent light from the fluorescent particles and calculated from a fluorescent image of a region identical to the region taken in the image from which the number of fluorescent particles is counted.