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Method for immunologically labeling tissue samples

A tissue sample and immune labeling technology, applied in the biological field, can solve problems that hinder technology promotion and application

Inactive Publication Date: 2016-12-07
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, when CLARITY technology is used to study the three-dimensional high-resolution structure of intact tissues, it takes a long time for labeled probes (especially antibodies) to reach the interior of the tissue from the surface of the intact tissue, which greatly hinders the promotion and application of this technology.

Method used

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  • Method for immunologically labeling tissue samples
  • Method for immunologically labeling tissue samples
  • Method for immunologically labeling tissue samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Example 1 Preparation of brain tissue samples from cleared mice

[0082] Degreased mouse brain slices were trimmed into rectangles with a razor blade and mounted between two coverslips. Adjust the thickness of the blue butadiene to ensure that the blue butadiene can seal the upper and lower edges of the brain slices after the brain slices are loaded. Between the two coverslips, add a small amount of epoxy glue along the edge of the blu-tack.

Embodiment 2

[0083] Example 2 Configuring an electric field device for clearing mouse brain tissue samples

[0084] Put the prepared glass slide-brain slice combination into a petri dish, drill two small holes with a diameter of 1 mm on the lid of the 35 mm petri dish, and place two platinum electrodes with a diameter of 0.5 mm and a length of 8 cm (purchased from Sigma ) through the small hole, bend the electrode into a right angle, reinforce the electrode at the small hole with blue butadiene glue, add 2ml antibody diluent to the petri dish, put the lid on the petri dish, and load the petri dish on On the microscope stage, clamp the electrode with alligator clips and connect it to the power supply (KXN-6020D, ZHAOXIN).

Embodiment 3

[0085] Example 3 Detection results of electric field accelerated antibody entry into brain tissue samples

[0086] experimental method:

[0087] 1) Dilute 6 μl of IgG (purchased from Molecular Probes Inc.) into 2000 μl of 0.1 M sodium borate buffer (pH 8.5).

[0088] 2) Add the antibody dilution to the culture dish, and cover the electrode cover.

[0089] 3) After finding the diffusion edge of the sample under the microscope, take pictures and record it.

[0090] 4) After standing still for 30 minutes, take pictures again to record the diffusion of antibody molecules in the brain slices.

[0091] 5) Turn on the power, adjust the voltage to 25V, take pictures and record after electrophoresis for 30 minutes.

[0092] 6) Process pictures and extract information.

[0093] Experimental results:

[0094] (1) if Figure 1-2 As shown, the results showed that under the action of the electric field, the antibody filled the whole brain slice more uniformly. Compared with the contro...

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Abstract

The invention discloses a method for immunologically labeling tissue samples, and particularly provides a labeling system. The labeling system comprises the tissue samples to be immunologically labeled, a probe and buffer solution. The probe and the buffer solution are used for labeling the tissue samples; labeling treatment can be carried out by the labeling system under the effects of electric fields, so that the probe can be fed into the tissue samples, the tissue samples can be immunologically labeled, and immunologically labeled tissue samples can be obtained. The method has the advantages that the duration in which the probe is fed into the tissue samples can be shortened, the internal integrity of tissues can be kept, and accordingly the method has great application value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for immunolabeling tissue samples. Background technique [0002] Studying the three-dimensional spatial structure of biomedical tissues at the cellular and subcellular scales is the basis for understanding their normal functions, and can also provide a basis for understanding the occurrence and development of organ diseases. Previous research on human and other animal tissues was mainly at the anatomical scale, while research at the cellular and subcellular scales was limited by the ability to analyze, and usually only the structural information of tissue slices could be studied. It is very time-consuming and labor-intensive to study tissue based on the 3D reconstruction technology of serial tissue slices. [0003] In recent years, the rapid development of tissue clearing technology has made it possible for people to obtain high-resolution three-dimensional stru...

Claims

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Application Information

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IPC IPC(8): G01N33/58
Inventor 邵志峰李小卫李俊丹尼尔·恰科夫
Owner SHANGHAI JIAO TONG UNIV