Catalytic De-Crosslinking for In Situ Analysis of Fixed Samples
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Solution Overview
Problem
Current methods for analyzing analytes in biological samples, such as nucleic acids and proteins, suffer from low sensitivity and specificity due to background autofluorescence and molecular crosslinks that render analytes less accessible, leading to biased and labor-intensive processes.
Innovation Solution
A method involving a catalyst that catalyzes the de-crosslinking of molecular crosslinks in fixed biological samples, followed by contacting with labelling agents and detecting optical signals to enhance analyte detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If formalin-fixation is used to preserve biological samples, then sample stability is improved, but background autofluorescence increases and analyte accessibility decreases
Solution Approach 1:
The patent applies preliminary de-crosslinking treatment to fixed biological samples before performing in situ analysis. This preliminary action removes molecular crosslinks introduced during formalin-fixation, thereby reducing background autofluorescence and improving analyte accessibility while preserving sample stability for subsequent detection procedures
2Stability of the object's composition
If molecular crosslinks are present in fixed samples, then sample preservation is improved, but analyte accessibility to detection reagents decreases
Solution Approach 1:
The patent performs preliminary de-crosslinking treatment on fixed biological samples before applying detection reagents. This preliminary step breaks down molecular crosslinks that hinder reagent access to analytes, thereby improving analyte accessibility while maintaining sample preservation for accurate detection
3Device complexity
If conventional analysis methods are used on fixed samples, then procedural simplicity is maintained, but detection sensitivity and specificity decrease
Solution Approach 1:
The patent introduces a preliminary de-crosslinking step in the analysis workflow for fixed biological samples. This additional step improves detection sensitivity and specificity by removing interfering crosslinks and reducing autofluorescence, while maintaining overall procedural simplicity through standardized treatment protocols
Solution Approach 2:
The patent employs de-crosslinking agents as intermediary substances that temporarily interact with fixed samples to remove crosslinks. These intermediaries facilitate improved analyte accessibility and reduced autofluorescence without permanently altering sample structure, enabling subsequent detection with enhanced sensitivity and specificity
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
Improves analyte detection sensitivity and specificity by reducing background autofluorescence and increasing the accessibility of molecular targets, allowing for more efficient and accurate analysis of nucleic acids and proteins in fixed tissues.
Implementation Method 1
contacting the biological sample with a catalyst that catalyzes de-crosslinking of molecular crosslinks in the biological sample
Data Source
AI summary
The present disclosure relates in some aspects to methods and compositions for in situ analysis involving catalytic de-crosslinking of biological samples.