Fluorescence Quantification Using Reference Substance Labeling
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Solution Overview
Problem
Current methods for observing multiple kinds of observation-target substances in a single tissue section using fluorescent dyes face challenges in separating excitation and emission wavelengths, making it difficult to accurately compare fluorescence intensity across different tissue sections and requiring multiple labeling substances, which limits the number of substances that can be effectively observed.
Innovation Solution
A quantification method that uses a reference substance labeled with one type of fluorescent dye and multiple quantification-target substances labeled with different fluorescent dyes, allowing for relative quantification without the need for multiple types of labeling substances, even when staining conditions vary across sections.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If multiple kinds of fluorescent dyes are used to observe multiple observation-target substances, then the number of observable substances increases, but the device complexity and difficulty of wavelength separation increase
Solution Approach 1:
The patent applies universality by using a single fluorescent dye that can label multiple different observation-target substances. Instead of requiring separate dyes for each target substance, the invention enables one fluorescent dye to serve multiple labeling functions through the use of multiple labeling substances (e.g., different antibodies or probes) that all emit the same fluorescence signal, thereby reducing device complexity while maintaining the ability to observe multiple substances
Solution Approach 2:
The patent employs copying by creating multiple identical fluorescent signals for different target substances. Each observation-target substance is labeled with a labeling substance that produces the same fluorescent output, effectively copying the fluorescence signal across multiple targets. This approach simplifies the detection system since only one fluorescence wavelength needs to be detected, while still allowing differentiation of multiple substances through their respective labeling substances
2Quantity of substance
If multiple tissue sections are stained to observe multiple observation-target substances, then the number of observable substances increases, but the measurement precision decreases due to inability to equalize staining conditions
Solution Approach 1:
The patent applies merging by combining multiple observation-target substances into a single tissue section for simultaneous observation. Instead of separating targets across multiple tissue sections, the invention enables co-labeling of multiple substances in one section using different labeling substances that all produce the same fluorescent signal, thereby maintaining measurement precision while increasing the number of observable substances
Solution Approach 2:
The patent applies local quality by allowing different labeling substances to be applied to different regions or targets within the same tissue section. Each labeling substance can be optimized for its specific target while all contributing to the same fluorescent channel, enabling both localized specificity and global comparability of fluorescence intensities across different targets in the same section
3Quantity of substance
If three or more kinds of labeling substances are used to label three or more observation-target substances, then the quantity of observable substances increases, but the ease of operation decreases due to filter selection requirements
Solution Approach 1:
The patent applies universality by designing a system where a single fluorescent dye and corresponding filter set can detect multiple different labeling substances. Instead of requiring unique filters for each fluorescent dye, the invention enables one filter configuration to universally detect all labeling substances, significantly simplifying the operational complexity while maintaining the capability to observe multiple substances
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 the accurate measurement of multiple observation-target substances using fewer types of fluorescent dyes, overcoming the limitations of existing methods by allowing for the observation and quantification of three or more substances without requiring multiple labeling substances, even when staining conditions differ.
Implementation Method 1
a first labeling substance that labels a reference substance commonly contained in the plurality of samples... detection value derived from a first labeling substance
Data Source
AI summary
A quantification method in the present invention is a quantification method for quantifying three or more kinds of quantification-target substances contained in a plurality of samples collected from one specimen, and includes a quantification step of quantifying the three or more kinds of quantification-target substances in each of the plurality of samples on the basis of a detection value derived from a first labeling substance that labels a reference substance commonly contained in the plurality of samples. In the present invention, three or more kinds of observation-target substances can be observed or quantified without requiring many kinds of labeling substances even when staining conditions are different from each other. That is, the observation-target substances (a reference substance and quantification-target substances other than the reference substance) can be each observed or quantified using a smaller number of kinds of labeling substances than the number of kinds of the observation-target substances.