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Tissue optical clearing agent and application thereof

A light-transparent agent and tissue technology, applied in the field of bio-optics, can solve the problems of affecting the second harmonic imaging of collagen, the destruction of collagen space structure, and the complex composition of light-transparent agent, so as to achieve uniform refractive index and endogenous The effect of strong optical signal and short time-consuming

Active Publication Date: 2015-04-29
SHENZHEN INST OF ADVANCED TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although the existing technology provides a variety of tissue light transparent agents, these technologies are mainly based on the basic principle of collagen dissociation, which will lead to the destruction of the spatial structure of collagen and the weakening or even disappearance of endogenous signals, which greatly affects the optical properties of collagen. Effects of Second Harmonic Imaging
In addition, the optical transparent agent disclosed in the prior art and its application method are time-consuming; the composition of the optical transparent agent is relatively complicated

Method used

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  • Tissue optical clearing agent and application thereof
  • Tissue optical clearing agent and application thereof
  • Tissue optical clearing agent and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0036] SD rats (200g) were killed by cervical dislocation after intraperitoneal injection of 1% pentobarbital 2ml, the hairs of the rats were removed with depilatory cream, the skin of the rats was cleaned with normal saline, and the skin of the back was removed with scissors. The knife removes the subcutaneous fat. After the rat skin was cleaned with phosphate buffer, soaked in 30% fructose, 10% polyethylene glycol monooctylphenyl ether (Sigma-Aldrich reagent, n=9~10) and 25% In the light clear agent aqueous solution composed of N,N,N',N'-tetra(2-hydroxypropyl)ethylenediamine, the skin becomes transparent after 15-20 minutes. Flatten the transparent skin, and use a two-photon fluorescence microscope to obtain the second harmonic signal map at a skin depth of about 120 μm (where the excitation wavelength is 900nm and the receiving wavelength is 360-540nm), to obtain Figure 1A . Use a two-photon fluorescence microscope to obtain the fluorescence signal map of skin blood vesse...

Embodiment 2

[0040] SD rats (200g) were killed by cervical dislocation after intraperitoneal injection of 1% pentobarbital 2ml, carefully took out the eyeball completely with tweezers and scissors, washed the residual blood on the eyeball with phosphate buffer, and placed it in a 4% polysaccharide POM was fixed for 24 hours, and washed with phosphate buffer after taking it out. Soak the prepared eyeballs in 70% fructose, 2% polyethylene glycol monooctylphenyl ether (Sigma-Aldrich reagent, n=9~10) and 5% N,N,N' , in the aqueous solution of light clearing agent composed of N'-tetrakis (2-hydroxypropyl) ethylenediamine, the eyeball becomes transparent after 15 to 20 minutes, such as figure 2 shown, where figure 2 The middle left picture is the eyeball before treatment, and the right picture is the transparent eyeball after treatment. Use a two-photon fluorescence microscope to obtain the second harmonic signal map of the eyeball at different depths (where the excitation wavelength is 900n...

Embodiment 3

[0042] SD rats (200g) were killed by cervical dislocation after intraperitoneal injection of 1% pentobarbital 2ml, carefully took out the eyeball completely with tweezers and scissors, washed the residual blood on the eyeball with phosphate buffer, and placed it in a 4% polysaccharide POM was fixed for 24 hours, and washed with phosphate buffer after taking it out. Soak the prepared eyeballs in 50% fructose, 5% polyethylene glycol monooctylphenyl ether (Sigma-Aldrich reagent, n=9~10) and 10% N,N,N' ,In the aqueous solution of light-transparency agent composed of N'-tetra(2-hydroxypropyl)ethylenediamine, the eyeball becomes transparent after 15 to 20 minutes, and the second harmonic signal of the eyeball at a depth of 170 μm is obtained with a two-photon fluorescence microscope Fig. (wherein excitation wavelength is 900nm, receiving wavelength is 360~540nm), obtain Figure 4 . From Figure 4 It can be seen that the second harmonic signal of the eyeball at 170 μm is clearly v...

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Abstract

The invention provides a tissue optical clearing agent and an application thereof. The tissue optical clearing agent comprises 30-70 wt% of fructose, 2-10 wt% of octyl phenoxy poly ethoxy, 5-25 wt% of N,N,N',N'-tetra (2-hydroxypropyl) ethylene diamine and the balance of water. The tissue optical clearing agent does not damage the spatial structure of collagen in tissue, a secondary harmonic signal diagram or a fluorescent signal diagram of collagen of the tissue can be obtained, and the tissue optical clearing agent can greatly increase an imaging depth.

Description

technical field [0001] The invention relates to a tissue light-transparency agent and an application thereof, in particular to a tissue light-transparency agent capable of maintaining the spatial structure of tissue collagen and an application thereof, belonging to the field of bio-optics. Background technique [0002] With the development of modern optical imaging technology and the emergence of various labeling technologies, it is possible to obtain high-resolution information on the structure and function of biological tissues, but the high scattering characteristics of biological tissues seriously restrict the application of optical imaging technology in the field of biomedicine. Multiphoton microscopy imaging technology uses near-infrared light to excite chromophores to obtain fluorescence or harmonic signals to improve the imaging depth. However, for biological tissues with large scattering, the penetration depth is still very limited. [0003] The optical transparency...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N21/64
Inventor 郑炜张洋
Owner SHENZHEN INST OF ADVANCED TECH
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