Analyte Detection via Antigen Retrieval and Multiplexing
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Solution Overview
Problem
Current methods for detecting objects within samples, such as biological fluids, face challenges in efficiently and accurately identifying biomarkers and target analytes due to limitations in imaging and labeling techniques, which often result in reduced sensitivity and specificity.
Innovation Solution
A method involving antigen retrieval steps, multiplexing with various stains and detection moieties, and amplification techniques to enhance signal detection, allowing for the simultaneous imaging of multiple biomarkers and improved epitope detectability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional imaging and labeling techniques are used, then the detection process is simple, but the sensitivity and specificity of detecting biomarkers are reduced
Solution Approach 1:
The detection process is divided into multiple sequential steps: initial labeling with first detection reagents, imaging to identify target objects, antigen retrieval treatment, and secondary labeling with second detection reagents. This segmentation allows each step to be optimized independently, improving overall detection accuracy while managing complexity through structured progression.
Solution Approach 2:
The method performs preliminary labeling and imaging before antigen retrieval to identify which specific target objects require further analysis. This preliminary action allows the subsequent complex antigen retrieval and secondary labeling steps to be focused only on relevant samples, improving detection accuracy while reducing unnecessary complexity.
2Adaptability or versatility
If single-marker detection is used, then the analysis is simpler, but the ability to detect multiple biomarkers simultaneously is limited
Solution Approach 1:
The method employs multiple detection reagents (both first and second sets) that can bind to different biomarkers, allowing a single detection system to perform multiple functions. The antigen retrieval step serves as a universal preparation that enables subsequent detection of various epitopes, making the system adaptable to detect multiple biomarkers simultaneously.
Solution Approach 2:
The detection process uses nested labeling strategies where second detection reagents are applied after and build upon the initial labeling with first detection reagents. This nested approach allows multiple layers of detection to be combined, enabling simultaneous detection of multiple biomarkers while organizing the complexity in a structured, manageable manner.
3Measurement precision
If antigen retrieval is performed, then epitope detectability is improved, but the detection process time is increased
Solution Approach 1:
The method performs preliminary detection with first labeling reagents before antigen retrieval to identify target objects that require epitope enhancement. This allows the time-consuming antigen retrieval step to be applied selectively only to samples where it will improve epitope detectability, rather than to all samples, thus balancing precision improvement with time efficiency.
4Measurement precision
If signal amplification is used, then detection sensitivity is improved, but the complexity of the detection method is increased
Solution Approach 1:
The method extracts and focuses amplification signals through sequential detection rounds. By performing initial labeling, imaging to identify targets, then applying antigen retrieval and secondary labeling specifically to enhance signals from identified targets, the system extracts the essential amplification benefit while avoiding the complexity of continuous or indiscriminate signal amplification throughout the entire process.
Data Source
AI summary
This disclosure relates generally to detection and, in particular, to detecting objects within a sample or fraction thereof.