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Simple and rapid pig blastaea differential dyeing method

A differential dyeing, pig blastocyst technology, applied in the field of developmental biology, can solve the problems of long dyeing cycle and low dyeing efficiency, and achieve the effect of shortening the experimental time, simplifying the experimental steps, and reducing the experimental cost.

Inactive Publication Date: 2013-08-14
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing technology has defects such as low staining efficiency and long staining cycle for staining pig blastocyst cells

Method used

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  • Simple and rapid pig blastaea differential dyeing method
  • Simple and rapid pig blastaea differential dyeing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The composition of Liquid 1 was: PBS containing 30ug / mL PI and 0.02% (V / V) TritonX100.

[0020] The composition of liquid 2 is: PBS containing 10ug / mL Hoechst33342.

[0021] The present invention will be further described below in conjunction with the accompanying drawings.

[0022] Preparation:

[0023] 1. Press the button before the experiment figure 1 Instructions Add the corresponding liquid in one row of the 24-well plate. Each row is a group, and when multiple groups are required for simultaneous staining, add the liquid to the other rows of the 24 plate in sequence according to the order given in the first row. Each row is for a set of experiments.

[0024] 2. Add 5ul of anti-quencher to the slide for final compression.

[0025] 3. Draw two glass mouth straws with a diameter larger than that of the blastocyst, and burn the mouth to prevent the glass puncture from scratching the blastocyst during the operation.

[0026] 4. At the beginning of the experiment,...

Embodiment 2

[0038] The difference with embodiment 1 is:

[0039] Liquid 1 composition: The composition of liquid 1 is: PBS containing 10ug / mL PI and 0.1% (V / V) TritonX100.

[0040] The composition of liquid 2 is: PBS containing 20ug / mL Hoechst33342.

Embodiment 3

[0042] The difference with embodiment 1 is:

[0043] Liquid 1 composition: The composition of liquid 1 is: PBS containing 50ug / mL PI and 0.01% (V / V) TritonX100.

[0044] The composition of liquid 2 is: PBS containing 20ug / mL Hoechst33342.

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Abstract

The invention discloses a simple and rapid pig blastaea differential dyeing method and belongs to the technical field of developmental biology. The method comprises the following steps: performing surfactant TritonX100 permeable membrane treatment on the pig blastaea, and performing differential dyeing on trophoderm cells on the outer layer of the blastaea and inner cell mass cells by utilizing the difference of two nucleus dyes in molecular size. According to the method, the experimental steps of performing differential dyeing on the pig blastaea are simplified, the experimental time is greatly shortened, and the experimental cost is reduced.

Description

technical field [0001] The invention relates to a simple and rapid differential staining method for pig blastocysts, which belongs to the technical field of developmental biology. Background technique [0002] Pig is not only an important meat animal, but also a good potential biomedical model animal. However, with the progress of pig-related research topics, more and more scholars have realized that the analysis of early embryonic development in pigs is far less clear than that in mice, which has constituted a bottleneck in the development of pig biomedical models. The application of fluorescent dyes in cell counting can be traced back to the 1980s, the first attempt by Ebert et al. in rodents and the subsequent use in other mammalian studies. The prior art has defects such as low staining efficiency and long staining cycle for staining pig blastocyst cells. Contents of the invention [0003] The invention provides a simple and fast differential staining method for pig ...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N21/64
Inventor 格日乐其木格刘忠华何文腾朱江
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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