Fluorescent Particle Quantification via Luminance Correction
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Solution Overview
Problem
Existing methods for quantifying biological substances in tissue samples using fluorescent dye accumulated particles face challenges due to luminance variations and variations in the number of particles adhering to the sample, which affect accuracy in pathological diagnosis.
Innovation Solution
A biological substance quantification method that involves inputting fluorescent images from a target sample and standard samples, calculating luminance values, and using these values to correct the number of fluorescent dye accumulated particles, including steps for unit luminance value calculation, tentative particle number calculation, and particle number correction to account for luminance variations and particle adherence.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If fluorescent bright spots are extracted and counted from fluorescent images to quantify biological substances, then the quantification process is simple, but the accuracy is low because multiple particles may appear as a single bright spot and luminance variations affect measurement precision
Solution Approach 1:
The patent segments the quantification process into multiple stages: first extracting bright spots, then performing luminance correction using reference images, and finally calculating particle numbers. This segmentation allows each stage to address specific problems independently, improving overall accuracy while maintaining operational simplicity.
Solution Approach 2:
The patent introduces reference images (third fluorescent images with known particle numbers) as an intermediary element to correct luminance variations in the target sample images. This intermediary enables the system to compensate for luminance differences caused by various factors without complicating the basic quantification workflow.
2Measurement precision
If luminance correction methods such as sensitivity calibration or standardization are applied to correct luminance variations, then measurement precision improves, but device complexity increases due to additional calibration procedures
Solution Approach 1:
The patent creates reference images that copy the staining conditions and luminance characteristics of the target samples. By using these copied reference images for correction, the system avoids the need for separate sensitivity calibration procedures while achieving similar accuracy improvements.
Solution Approach 2:
The patent performs luminance correction using pre-acquired reference images before the actual quantification measurement. This preliminary action eliminates the need for on-demand calibration procedures, reducing device complexity while maintaining measurement precision.
3Productivity
If the number of fluorescent dye accumulated particles is calculated based on average luminance value per particle, then quantification can be performed, but accuracy deteriorates due to luminance variations from different staining conditions, light source intensity, and microscope differences
Solution Approach 1:
The patent implements a feedback mechanism where reference images with known particle numbers are used to establish luminance correction factors. These correction factors are then applied to the target samples to compensate for luminance variations, enabling accurate and rapid quantification without manual calibration.
Solution Approach 2:
The patent changes the luminance parameter by applying correction factors derived from reference images to the target sample images. This parameter transformation compensates for variations in staining conditions, light source intensity, and microscope differences, improving calculation accuracy while maintaining productivity.
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 accurate quantitative analysis of biological substances by correcting for luminance variations and particle adherence, improving the reliability of pathological diagnosis.
Implementation Method 1
fluorescent dye accumulated particle bondable to a specific biological substance... image capturing of the target sample... fluorescent image
Data Source
AI summary
A biological substance quantification method of quantifying an expression amount of a specific biological substance in a target sample stained by using a fluorescent dye accumulated particle bondable to the specific biological substance. The method includes: inputting a first fluorescent image obtained by image capturing of the target sample; extracting a bright spot portion from the first fluorescent image and calculating a first luminance value; and calculating a number of the fluorescent dye accumulated particle included in the bright spot portion by using the first luminance value, a second luminance value of a bright spot portion extracted from a second fluorescent image obtained by image capturing of a standard sample, and a distribution of a third luminance value of each bright spot portion in a third fluorescent image obtained by image capturing of a preparation on which the fluorescent dye accumulated particle is dispersed without aggregating.