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A tissue light transparent agent and its preparation method and application

A light transparent agent and tissue technology, applied in the field of biomedicine and transparent sample preparation, can solve the problems of changing the shape of the sample, disappearing of the natural texture of the tissue, and the quenching of the fluorescent marker signal of the tissue, so as to achieve the improvement of imaging depth and quality, depth and signal The effect that the noise ratio is improved and the optical intensity is not affected

Active Publication Date: 2019-10-11
SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The tissue transparent technologies used in the prior art include BABB technology, Scale technology, SeeDB technology, CLARITY technology, etc. Among them, the BABB technology uses organic solvents. Although the tissue transparent effect is good after treatment, the tissue shrinks severely when it is processed, and causes fluorescence in the tissue. The marker signal is quenched; the Scale technology is to continuously soak the biological tissue in the urea solution, which takes a long time to make the tissue transparent, and will also cause serious swelling of the tissue; the SeeDB technology uses the fructose solution with increasing concentration to soak the tissue, and the tissue shape Although it is well maintained, the tissue often turns brown, and the fructose solution is viscous, which makes it inconvenient to operate; CLARITY technology uses electrophoresis to remove the lipids in the tissue that cause scattering to achieve transparency. This method has a good effect on tissue transparency, but it needs to be The tissue is pre-fixed with a special hydrogel, and the fat-dissolving will cause the destruction of the cell membrane and the disappearance of the natural texture of the tissue
[0004] The tissue light transparency schemes reported above will more or less change the shape of the sample, destroy the natural texture structure of the tissue, damage the optical signal, and affect the imaging results

Method used

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  • A tissue light transparent agent and its preparation method and application
  • A tissue light transparent agent and its preparation method and application
  • A tissue light transparent agent and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] A preparation method of a tissue light transparent agent, comprising the following steps:

[0043] Mix urea, formamide, Triton X-100 (Sigma-Aldrich reagent), N,N,N',N'-tetrakis(2-hydroxypropyl)ethylenediamine with water and heat at 35°C Dissolve until the solution is clear and transparent to obtain the tissue light transparent agent, wherein the mass fraction of each component in the tissue light transparent agent is: urea: 25%, formamide: 10%, triton: 20% , N,N,N',N'-tetrakis(2-hydroxypropyl)ethylenediamine: 25%, water: 20%. The tissue light transparent agent is suitable for the brain tissue of young mice, and the transparent effect is better when the whole brain tissue is transparent, and the tissue shape remains basically unchanged. Tissue transparency is achieved in 5-8min.

Embodiment 2

[0045] A preparation method of a tissue light transparent agent, comprising the following steps:

[0046] Mix urea, formamide, Triton X-100, N,N,N',N'-tetrakis(2-hydroxypropyl)ethylenediamine with water, and heat to dissolve at 37°C until the solution is clear and transparent , to obtain the tissue light-transparent agent, wherein, the mass fraction of each component in the tissue light-transparent agent is: urea: 20%, formamide: 15%, triton: 10%, N, N, N ', N'-Tetrakis(2-hydroxypropyl)ethylenediamine: 20%, the balance is water. This tissue light clearing agent is suitable for tissues with high water content in young mice, such as lung tissue. Also suitable for thin slices of brain tissue. When transparent to thin slices of brain tissue, the transparent speed is relatively slow, but the final transparent effect is still ideal. During the transparent treatment, the transparent temperature of the transparent agent can be increased appropriately, and the transparent effect can...

Embodiment 3

[0048] A preparation method of a tissue light transparent agent, comprising the following steps:

[0049] Mix urea, formamide, Triton X-100, N,N,N',N'-tetrakis(2-hydroxypropyl)ethylenediamine with water, and heat to dissolve at 35°C until the solution is clear and transparent , to obtain the tissue light transparent agent, wherein, the mass fraction of each component in the tissue light transparent agent is: urea: 25%, formamide: 8%, triton: 2015%, N, N, N ', N'-Tetrakis(2-hydroxypropyl)ethylenediamine: 15%, the balance is water. The tissue light transparent agent is more suitable for the transparency of liver and kidney tissues, and the transparent time of the whole tissue can be controlled within 3-4 days.

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Abstract

The invention provides a tissue optical clearing agent which comprises, in weight percent, 15-25% of urea, 8-15% of formamide, 10-20% of triton, 20-25% of N,N,N',N'-Four (2-hydroxy propyl) ethylenediamine and the rest water. According to the tissue optical clearing agent, biological tissue scattering can be effectively reduced in a short time, tissue transparency is achieved, the integrity of biological tissues can be effectively kept, the volume of the biological tissues is basically unchanged, expansion or contraction is avoided, tissues cannot become brittle, so that imaging depth and signal-to-noise ratio are greatly improved, and endogenous fluorescent signal strength is basically unaffected. The invention further provides a preparation method and an application of the tissue optical clearing agent.

Description

technical field [0001] The invention relates to the technical field of biomedicine and transparent sample preparation, in particular to a tissue light transparent agent and its preparation method and application. Background technique [0002] With the continuous development of optical imaging technology, it has become possible to obtain high-resolution structure and function information of biological tissue, but the opacity of biological tissue restricts the application of optical imaging technology in the field of biomedicine. [0003] The tissue light-transparency technology that has emerged in recent years allows organs to be observed and studied in a complete state without slicing. The core of this technology is that the introduction of hypertonic and high-refractive chemical reagents into biological tissues can effectively reduce the scattering characteristics of biological tissues. Make the sample transparent, greatly improving the depth of imaging. The tissue transpa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/30
Inventor 郑炜夏先园张洋
Owner SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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