Biotissue Clearing Composition Using CHAPS and Urea
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for clearing biotissue are costly, time-consuming, and inefficient, particularly for obtaining three-dimensional images at the cellular level, as they require expensive equipment and solvents, and often result in tissue hardening, reduced antibody permeability, and loss of antigenicity, limiting their application to various tissues.
Innovation Solution
A composition comprising CHAPS and urea, used in a clarity method that contacts fixed biotissue to remove lipids without causing structural damage, allowing deep light penetration and improving antibody staining, applicable to various tissues including brain, liver, and muscle, without the need for high-cost electrophoresis apparatus or additional refractive index adjustment solutions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional clarity methods using organic solvents or polymers are used, then tissue clearing is achieved, but fluorescence and antigen conservation decrease
Solution Approach 1:
The invention changes the chemical parameters of the clearing solution by using a specific composition (30-70% v/v TCA, 10-30% v/v aceton, 5-20% v/v methanol, 5-30% v/v water) instead of conventional organic solvents or polymers. This parameter change enables effective lipid removal while preserving fluorescent proteins and antigenicity, resolving the contradiction between clearing effectiveness and substance conservation.
2Strength
If hydrogel concentration is increased to improve tissue support, then binding with protein increases and net structure gets tighter, but tissue becomes harder and lipid escape becomes difficult, extending clearing time
Solution Approach 1:
The invention extracts and eliminates the hydrogel polymer component from the clearing system entirely. By using only the chemical clearing solution (TCA-aceton-methanol-water composition) without any hydrogel supporter, the method avoids the contradiction between providing tissue support and enabling lipid escape, achieving both effective clearing and preserved tissue integrity.
3Reliability
If conventional clarity methods are used, then tissue clearing is achieved, but air and dark particles are deposited on tissue surface, causing yellow discoloration
Solution Approach 1:
The invention changes the chemical composition parameters by using a specific TCA-aceton-methanol-water formula that prevents particle deposition and yellow discoloration. This parameter optimization resolves the contradiction between achieving effective clearing and avoiding harmful side effects like particle deposition and tissue discoloration.
4Strength
If polyacrylamide net structure is used to support tissue, then tissue integrity is maintained, but antibody staining material diffusion is significantly hindered
Solution Approach 1:
The invention removes the polyacrylamide net structure from the system entirely. By using only the chemical clearing solution without any polymer network, the method eliminates the barrier to antibody diffusion while still maintaining tissue integrity through the clearing process itself, resolving the contradiction between structural support and reagent penetration.
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 effectively clears biotissue, preventing bubble formation and color changes, enhancing tissue clarity, and facilitating antibody-staining, enabling three-dimensional observation and disease diagnosis without damaging tissues, thus improving the efficiency and cost-effectiveness of tissue analysis.
Implementation Method 1
uses a net supporter which is constructed in a tissue to hold materials important for diagnosis such as DNA or proteins by using hydrogel and eliminates lipids selectively
Implementation Method 2
The composition for clearing biotissue comprising the compound represented by formula 1 below, the optical isomer, the hydrate, or the salt of the same
Data Source
AI summary
The present invention relates to a composition for clearing biotissue and a clarity method for biotissue using the same. The composition for clearing biotissue and the clarity method for biotissue using the same of the present invention do not need a high price electrophoresis apparatus and expensive solutions, and can not only be applied to various biotissues including brain, liver, lung, kidney, intestine, heart, muscle, and blood vessel, without damaging any of them but also prevent bubble formation, color change, and dark sediment; improve the clarity of biotissues; and help antibody-staining of the cleared tissues. Therefore, it is useful to identify the reason of various diseases through structural imaging of biotissue and to establish a treatment method.