Dual-Staining Tissue Analysis for Quantitative Pathology
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Solution Overview
Problem
Conventional staining methods for tissue samples face challenges in achieving precise and quantitative evaluation of biological substance expression and localization, with bright field observation methods lacking quantitativeness and light-emitting substance methods lacking detailed pathology judgment.
Innovation Solution
A method that combines bright field observation and fluorescence staining on the same tissue section using labeled probe biological substances with pigmentation-inducing and fluorescent labels, allowing for detailed and quantitative assessment of biological substance expression and localization.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of information
If a staining method using enzyme reaction (DAB method) is used to allow bright field observation, then detailed information on the target molecule can be obtained through comprehensive judgment of color and staining characteristics, but the stainability changes depending on staining conditions including reaction time, resulting in poor quantitativity
Solution Approach 1:
The patent combines two different staining methods (bright field observation using DAB and fluorescence observation) into a single dual-staining procedure. This allows simultaneous acquisition of detailed morphological information from bright field and quantitative data from fluorescence, resolving the contradiction between information richness and measurement precision
Solution Approach 2:
The patent introduces fluorescence intensity as a new measurable parameter that is less sensitive to staining condition variations. By measuring fluorescence intensity digitally, the system achieves quantitative precision while maintaining the detailed information capability of traditional bright field observation
2Measurement precision
If a visualization method using a light-emitting substance (phosphor) is used, then quantitative evaluation can be carried out with excellent reproducibility, but detailed pathology judgment based on accumulated know-how of skilled pathologists cannot be expected
Solution Approach 1:
The patent merges fluorescence staining (providing quantitative data) with bright field staining (providing detailed morphological information). This combination allows pathologists to utilize both the quantitative precision of fluorescence and the detailed visual assessment capabilities developed through experience
Solution Approach 2:
The dual-staining method creates a multi-functional observation system that serves both quantitative measurement and qualitative assessment purposes simultaneously, making the technique universally applicable for both research and diagnostic applications
3Extent of automation
If only fluorescence staining is performed, then digital analysis of location and expression amount can be achieved, but the staining process and evaluation criteria become more complex compared to conventional bright field methods
Solution Approach 1:
By combining fluorescence and bright field staining in one protocol, the patent enables automated digital analysis of fluorescence while simultaneously providing familiar bright field images for verification, thereby managing complexity through integration rather than replacement
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 high-precision, quantitative evaluation of biological substance expression and localization with both visual and digital analysis, reducing variability in pathology judgments and improving the effectiveness of pharmaceutical assessments.
Implementation Method 1
a method, in which an anti-HER2 antibody bound to a HER2 antigen site in a tissue sample is stained and visualized using diaminobenzidine (DAB)
Implementation Method 2
a technique that uses a substrate to be converted into a pigmentary substance by an enzyme
Implementation Method 3
a visualization method that uses a substance having a color-developing property in itself... phosphors are suitably used therein (immunostaining that uses a phosphor is hereinafter also referred to as 'immunofluorescence staining')
Data Source
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AI summary
Provided is a method for staining a tissue enabling highly precise staining, by which the expression amount and/or the location of a biological substance in a tissue sample can be detected with a high quantitativity together with detailed information that can be obtained by bright field observation. The tissue staining method of the present invention is a method for staining a tissue, in which both staining that allows bright field observation and fluorescence staining are carried out for the same specific biological substance.