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Animal tissue transparentizing method

An animal tissue and tissue technology, applied in the field of animal tissue observation, can solve the problems of protein observation, difficulty in imaging, tissue deformation, etc., and achieve the effects of shortened transparency time, high safety, and easy operation.

Pending Publication Date: 2020-10-02
SHANDONG UNIV OF TRADITIONAL CHINESE MEDICINE
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Problems solved by technology

[0003] Most of the currently used tissue clearing techniques are organic solvents or water-soluble solvents. After organic solvents are cleared, imaging is more difficult and will cause corrosion to the lens. Water solvents take a long time to clear and cause large tissue deformation, which may cause serious damage to the tissue. There is a certain impact on the later protein observation

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Embodiment 1

[0047] Reagents required for the experiment:

[0048] Anesthetic: 0.3% pentobarbital (0.3g pentobarbital dissolved in 100mL deionized water) and ketamine (90mg / kg);

[0049] Phosphate buffered saline solution (PBS): 1×PBS is 0.01MPBS, after dissolving 22.4g (phosphate buffered solution, PBS) buffer dry powder with 2L deionized water, adjust the pH value to 7.2-7.4.

[0050] Tissue fixation solution: 4% paraformaldehyde, that is, 4g of PFA (Paraformaldehyde, PFA) powder dissolved in 1×PBS to 100mL;

[0051] Cardiac perfusate: 0.9% normal saline injection or 0.05 MEDTA, that is, 0.05 mol ethylenediaminetetraacetic acid (EDTA) is dissolved in deionized aqueous solution to a total volume of 1 L;

[0052] 0.1% Triton solution: Put TritonX-100 in a water bath at 37°C to 40°C for 2 to 3 hours to fully dissolve and mix well, then take 100μL TritonX-100 and dissolve it in 1×PBS to 100mL.

[0053] 3% BSA solution: Dissolve 3g of BSA powder in 0.1% Triton solution to 100mL.

[0054] 5...

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Abstract

The invention provides an animal tissue transparentizing method, which belongs to the technical field of animal tissue observation, and comprises the following steps of: 1) anesthetizing an animal, and sequentially pouring heart perfusate and tissue fixing liquid into the heart of the animal; dissecting and separating animal tissues; 2) placing the bone tissue in a decalcification solution for decalcification for 5-12 days to obtain a decalcified bone tissue, wherein in the decalcification process, the liquid is changed once every 1-3 days; 3) respectively putting the decalcified bone tissue and soft tissue into a decolorizing solution for decolorizing to obtain a decolorized tissue; 4) placing the decolorized tissue in a degreasing liquid for gradient degreasing, and then placing the tissue in a dehydration liquid for dehydration to obtain a dehydrated tissue; and 5) placing the dehydrated tissue in a transparent liquid for transparentizing to obtain a transparent tissue. Through themethod, large-view, multi-angle and deep observation of the tissue can be realized.

Description

technical field [0001] The invention belongs to the technical field of animal tissue observation, in particular to a method for transparentizing animal tissue. Background technique [0002] Organizational observation is the most common observation method in medical and scientific research. Traditional histological methods mainly include gross dissection and tissue sectioning. Gross anatomy can observe the position, shape, structure and adjacency of various organs in animals, but it cannot observe the microstructure in the organs. Tissue sections are divided into paraffin-embedded sections, frozen sections, etc. Although it can obtain detailed information about cells or subcellular structures in organs through a microscope, it also has its limitations: first, the integrity of organs is destroyed during the tissue sectioning process; Continuity of microstructure. In the past few decades, with the rapid development of imaging technology, imaging methods such as CT, MRI, and...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N21/64
CPCG01N1/30G01N21/6458G01N21/6428G01N2001/305G01N2021/6439
Inventor 夏明峰相珊郑志娟阚东方李静李运伦
Owner SHANDONG UNIV OF TRADITIONAL CHINESE MEDICINE
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