Hydrophilic Ionic Liquid Formulation for Deformation-Free Tissue Clearing
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing biological tissue treatment methods face challenges such as tissue deformation, high toxicity, slow clearing speed, ice crystal formation, and low fluorescence intensity, which affect the effectiveness and safety of tissue clearing, cryopreservation, fluorescence enhancement, and super-resolution imaging.
Innovation Solution
A hydrophilic amorphous ionic liquid formulation comprising specific cations and anions, with optional auxiliary agents, is used for refractive index matching, cryopreservation, and fluorescence enhancement, maintaining a stable liquid state and preventing ice crystal formation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Illumination intensity
If organic solvents are used for tissue clearing, then refractive index matching and clearing effect are improved, but fluorescence preservation deteriorates and tissue distortion increases
Solution Approach 1:
The patent changes the chemical composition parameters by using ionic liquid components (choline chloride, urea, nicotinamide) instead of traditional organic solvents, achieving high refractive index (1.33-1.45) while maintaining fluorescence preservation through non-toxic, non-denaturing properties of the ionic liquid formulation
Solution Approach 2:
The patent creates a composite ionic liquid formulation combining multiple components (choline chloride, urea, nicotinamide, and water) to achieve synergistic effects: choline chloride provides high refractive index, urea prevents protein denaturation, and nicotinamide enhances fluorescence preservation, resolving the contradiction between clearing effectiveness and fluorescence preservation
2Reliability
If aqueous clearing methods are used, then fluorescence preservation is improved, but tissue expansion and rigidity loss occur
Solution Approach 1:
The patent adjusts the refractive index parameter of the aqueous-based ionic liquid formulation to match tissue refractive index (1.33-1.45) through controlled composition ratios, eliminating the need for tissue expansion while maintaining fluorescence preservation and providing rigidity support for sectioning
3Reliability
If high concentration cryoprotectant is used for vitrification, then ice crystal formation is prevented, but toxicity to tissue increases
Solution Approach 1:
The patent changes the cryoprotectant composition parameters by using ionic liquid components with different freezing points (choline chloride -30°C, urea -6.7°C, nicotinamide 128°C) to achieve vitrification at higher temperatures with lower effective concentration, reducing toxicity while preventing ice crystal formation through the amorphous solid state formation
Solution Approach 2:
The ionic liquid formulation acts as an intermediary substance that mediates between the need for ice crystal prevention and tissue toxicity reduction, forming an amorphous solid state that protects tissue structure without the extreme toxicity of traditional high-concentration cryoprotectants
4Reliability
If traditional cryopreservation methods are used, then ice crystal formation is reduced, but recrystallization during rewarming damages tissue
Solution Approach 1:
The patent changes the phase transition parameters by using ionic liquid components with eutectic properties that lower the freezing point and broaden the temperature range for amorphous solid formation, preventing recrystallization during rewarming by maintaining the amorphous structure through controlled composition ratios
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 ionic liquid achieves transparent tissue clearing without deformation, effective cryopreservation, enhanced fluorescence at low temperatures, and ice-free cryosectioning, enabling high-resolution imaging and long-term sample preservation.
Implementation Method 1
uses refractive index matching solution to match the refractive index of tissue to a higher uniform level, thereby reducing light scattering, refraction and reflection, and making the treated tissue show a transparent effect
Implementation Method 2
Cryopreservation of Tissue by Vitrification at Ultra-Low Temperature uses cryoprotectant with high concentration and liquid nitrogen to cool down, so that dehydrated samples can be vitrified without the production of ice
Implementation Method 3
the fluorescence intensity of the fluorescent substance at low temperature is significantly enhanced as compared with that at room temperature
Data Source
AI summary
A hydrophilic amorphous ionic liquid preparation and a preparation method therefor, a biological tissue processing method using the ionic liquid preparation, and an application of the ionic liquid preparation in biological tissue processing. The ionic liquid preparation comprises: a) an ionic liquid containing a specific cation and a specific anion; b) an auxiliary agent for adjusting the properties of the components of a); and c) water.