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Method of immunolabeling tissue sample

A tissue sample, immune labeling technology, applied in the biological field, can solve problems such as hindering the promotion and application of technology

Active Publication Date: 2020-08-25
上海诒福科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 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 of immunolabeling tissue sample
  • Method of immunolabeling tissue sample
  • Method of immunolabeling tissue sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Embodiment 1 prepares the brain tissue sample of clearing mouse

[0083] 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

[0084] Example 2 Configure an electric field device for the brain tissue samples of cleared mice

[0085] 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

[0086] Example 3 Detection Results of Antibody Accelerated by Electric Field into Brain Tissue Sample

[0087] experimental method:

[0088] 1) Dilute 6 μl of IgG (purchased from Molecular Probes Inc.) into 2000 μl of 0.1 M sodium borate buffer (pH 8.5).

[0089] 2) Add the antibody dilution to the culture dish, and cover the electrode cover.

[0090] 3) After finding the diffusion edge of the sample under the microscope, take pictures and record it.

[0091] 4) After standing still for 30 minutes, take pictures again to record the diffusion of antibody molecules in the brain slices.

[0092] 5) Turn on the power, adjust the voltage to 25V, take pictures and record after electrophoresis for 30 minutes.

[0093] 6) Process pictures and extract information.

[0094] Experimental results:

[0095] (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 control gr...

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Abstract

The invention discloses a method for immunolabeling a tissue sample, and particularly provides a labeling system which comprises the tissue sample to be immunolabeled, a probe for labeling the tissuesample and a buffer solution, and (b) labeling the labeling system under the action of an electric field, so that the probe enters the tissue sample, the tissue sample is subjected to immunolabeling,and an immunolabeled tissue sample is obtained. According to the invention, the time for the probe to enter the tissue sample can be shortened, the integrity of the interior of the tissue can be maintained, and the application value is great.

Description

[0001] This application is a divisional application of an invention patent application with an application date of April 30, 2015, an application number of 201510219570.4, and an invention title of "A Method for Immunolabeling Tissue Samples". technical field [0002] The invention belongs to the field of biotechnology, and in particular relates to a method for immunolabeling tissue samples. Background technique [0003] 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-intens...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533
Inventor 邵志峰李小卫李俊丹尼尔·恰科夫
Owner 上海诒福科技有限公司