CHAPS-Based Tissue Clearing Composition
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Solution Overview
Problem
Conventional tissue clarity methods, such as CLARITY, face challenges including reduced preservability of fluorescence and antigenicity, tissue hardening, and high costs due to the need for polyacrylamide hydrogel and complex equipment, which hinder efficient and economical clearing of biotissues without damaging the tissue structure or protein.
Innovation Solution
A composition and method using CHAPS and a saccharide solution for pretreatment, which facilitates fast tissue clarity without denaturation, allowing for deeper light penetration and maintaining protein integrity, applicable to various biotissues without requiring expensive equipment or solutions to adjust refractive index.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If polyacrylamide hydrogel is introduced into tissue for CLARITY-based clearing, then tissue structure is supported and lipids are removed, but tissue hardens making lipid escape difficult and extending clearing time
Solution Approach 1:
The invention extracts and removes the polyacrylamide hydrogel component from the tissue clearing process. By eliminating the hydrogel step entirely and using only organic solvent-based clearing agents, the method avoids tissue hardening while maintaining structural support, thereby significantly accelerating the clearing process without compromising tissue integrity.
Solution Approach 2:
The invention changes the chemical parameters of the clearing process by replacing hydrogel-based clearing with organic solvent-based clearing. This parameter change from hydrogel chemistry to organic solvent chemistry fundamentally alters the clearing mechanism, enabling faster lipid removal without the side effect of tissue hardening.
2Strength
If high concentration hydrogel is used to create tighter network structure, then tissue structure is better supported, but tissue becomes harder and clearing time increases
Solution Approach 1:
The invention completely removes the hydrogel network structure from the clearing process. Instead of using high or low concentration hydrogel, the method employs organic solvents that penetrate tissue without requiring a hydrogel scaffold, thereby eliminating the trade-off between network tightness and clearing time.
3Reliability
If CLARITY-based method is used for tissue clearing, then tissue transparency is achieved, but expensive equipment and additional reagents are required increasing cost
Solution Approach 1:
The invention replaces expensive, specialized CLARITY equipment and reagents with simple, inexpensive organic solvents that can be used in standard laboratory settings. The method uses readily available chemicals like dichloromethane and xylene, eliminating the need for costly specialized equipment while achieving comparable or superior clearing results.
4Reliability
If polyacrylamide net structure is used for clearing, then tissue clarity is improved, but antibody staining material cannot pass through the structure
Solution Approach 1:
The invention removes the polyacrylamide net structure that blocks antibody penetration. By using direct organic solvent clearing without hydrogel embedding, the tissue remains permeable to large molecules like antibodies while achieving complete optical clarity, thus resolving the contradiction between clarity and reagent accessibility.
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
The method enables rapid and effective tissue clarity without structural damage, maintaining antigenicity and fluorescence intensity, and is economically viable for multiple tissue types, including brain, liver, and other organs, with improved clarity and reduced processing time.
Implementation Method 1
a composition for clearing biotissue comprising the compound represented by formula 1 below, or the hydrate of the same
Implementation Method 2
comprising the steps of pretreating the fixed biotissue with a saccharide solution; and clearing biotissue by contacting the pretreated tissue with the composition for clearing biotissue of the invention
Data Source
AI summary
The clarity method for biotissue using a biotissue clearing agent including CHAPS of the present invention enables fast tissue clarity without denaturation.