In Situ Immunofluorescence Quantification of Protein Expression
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Solution Overview
Problem
Current immunohistochemistry (IHC) methods struggle to provide continuous and precise quantification of protein expression in tumor tissues, leading to ambiguous results and loss of information, especially in cancer diagnosis, where precise molecular profiling is crucial for personalized treatments.
Innovation Solution
A method for quantitative measurement of target markers by in situ immunofluorescence, involving specific staining and image processing steps to establish a linear relationship between signal intensity and antigen expression, allowing for continuous scoring comparable to fluorescence in situ hybridization (FISH) and providing additional diagnostic information.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If chromogenic IHC staining is used to visualize immune complexes in tissue architecture, then detection of target presence is achieved, but continuous quantification of protein expression levels is lost
Solution Approach 1:
The patent replaces the enzymatic chromogenic detection system with a fluorescent imaging system. Instead of using enzyme-substrate reactions that produce colored precipitates (mechanical/chemical system), the invention uses fluorescently labeled antibodies detected by fluorescence microscopy (optical system), enabling continuous quantitative measurement of protein expression levels while preserving tissue morphology
Solution Approach 2:
The patent changes the detection parameter from binary chromogenic presence/absence to continuous fluorescence intensity measurement. By measuring fluorescence signal intensity rather than merely detecting presence of color, the system transforms the output from qualitative to quantitative, allowing precise measurement of protein expression levels across a continuous scale
2Ease of operation
If semi-quantitative scoring methods are used to assess biomarker expression, then diagnostic assessment is simplified, but continuous quantification and subtle expression differences are lost
Solution Approach 1:
The patent replaces manual semi-quantitative scoring by pathologists with automated fluorescence intensity measurement and image analysis. The system objectively measures fluorescence signal strength and correlates it with protein expression levels, eliminating subjectivity and rounding errors inherent in manual scoring while maintaining operational simplicity through automation
Solution Approach 2:
The patent creates a digital copy of the tissue expression pattern through fluorescence imaging and image analysis. Instead of relying on human interpretation of visual staining patterns, the system captures digital images and extracts quantitative data, preserving continuous expression information that can be precisely measured and compared without loss of subtle differences
3Measurement precision
If expensive complementary gene analysis methods are used to achieve precise molecular profiling, then accurate diagnosis is improved, but cost and complexity increase
Solution Approach 1:
The patent makes fluorescence in situ hybridization (FISH) methodology universal for both gene copy number detection and protein expression quantification. By applying the same fluorescent imaging platform to detect different targets (genes and proteins), the system eliminates the need for separate expensive gene analysis methods, reducing overall assay complexity while maintaining precise molecular profiling capabilities
Solution Approach 2:
The patent merges protein expression analysis and gene copy number detection into a single unified fluorescent imaging workflow. Instead of performing separate chromogenic IHC and FISH assays, the invention combines both detections using fluorescently labeled probes and antibodies detected by the same imaging system, simplifying the overall diagnostic process while providing comprehensive molecular profiling
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 and continuous quantification of protein expression levels, reducing the need for expensive complementary gene analysis and improving the precision of diagnosis and prognosis in cancer therapy.
Implementation Method 1
Method for quantitative measurement of a biomarker by in situ immunofluorescence
Implementation Method 2
incubating the sample with at least one target probe
Data Source
AI summary
The invention relates to a method for quantitative measurement of a biomarker by in situ immunofluorescence and uses thereof. In particular, the invention relates to a method which is a useful tool for use in the field of diagnosis, prevention and/or treatment of disease or disorders, in particular in the field of cancer management and therapy.