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Method for keeping integrity of embryos in early embryo tissue immunohistochemistry

A technology of immunohistochemistry and integrity, which is applied in the preparation, sampling, and measuring devices of test samples, which can solve the problems of easy adhesion of embryos and rupture of cyst wall, and achieve simple operation, improved effect, and strong pertinence Effect

Inactive Publication Date: 2018-06-29
ANHUI SCI & TECH UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, during the culture process, the embryos are easy to adhere to the wall, and the capsule wall will be broken when the culture medium is changed to move the embryos, so many embryos will be lost during the whole culture process.

Method used

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  • Method for keeping integrity of embryos in early embryo tissue immunohistochemistry

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Embodiment Construction

[0023] The methods used in the following examples are conventional methods unless otherwise specified, and the percentages are volume percentages unless otherwise specified.

[0024] 1. Implementation materials

[0025] Early parthenogenetic embryos constructed in vitro, including precleavage, 2-cell embryos, 8-cell embryos and 16-cell embryos.

[0026] 2. Implementation method

[0027] 1) Solution preparation: 4% paraformaldehyde, 0.5% Triton X-100, hydrogen peroxide 20% polyvinylpyrrolidone storage solution and 0.1μg / mL 4',6-diamidino-2-phenylindole all use the pH range 7.2-7.4 0.01mol / L phosphate buffer dilution; and immunohistochemistry with 4% paraformaldehyde, 0.5% Triton X-100, hydrogen peroxide and phosphate buffer were added to polyvinylpyrrolidone stock solution, the added concentration 2%, 4%, 6%, 8%, 10% respectively;

[0028] 2) Embryo washing: Under a microscope, pick out the parthenogenetic embryos, androgenetic embryos or in vitro fertilized embryos cultivat...

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Abstract

The invention discloses a method for keeping the integrity of embryos in early embryo tissue immunohistochemistry, and belongs to the technical field of animals and organisms. A polyvinylpyrrolidone solution is added to a phosphate buffer solution, paraformaldehyde, Triton X-100 and hydrogen peroxide which are used for the early embryo tissue immunohistochemistry to prevent embryo attachment, so the integrity of the embryos is kept in the whole immunohistochemistry process, and the immunohistochemistry effect is improved. The method mainly aims at in vitro embryos, and the components of a tissue treatment reagent are adjusted on the basis of adjusting routine immunohistochemistry steps, so the immunohistochemistry effect is effectively improved. The method has the advantages of overcomingthe disadvantages in the immunohistochemistry technology, enlargement of the application range, simplicity in operation, and low production cost.

Description

technical field [0001] The invention relates to a method for maintaining embryo integrity in early embryo tissue immunohistochemistry, and belongs to the field of animal biotechnology. Background technique [0002] Polyvinylpyrrolidone, referred to as PVP, is a polymer compound produced by the polymerization of N-vinyl-2-pyrrolidone. It is a white hygroscopic powder, odorless or slightly odorous, soluble in water, ethanol, chloroform and most organic solvents, insoluble in ether, and less toxic. Polyvinylpyrrolidone can be synthesized by monomer vinylpyrrolidone through bulk polymerization, solution polymerization and other methods. As a synthetic water-soluble polymer compound, it has the general properties of water-soluble polymer compounds, colloidal protection, film-forming, cohesive, hygroscopic, solubilizing or coagulating effects. Its solution has very low toxicity and is non-physiologically descriptive. It is widely used in medicine, food, cosmetics and other fiel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533G01N1/30G01N1/34G01N1/28
CPCG01N1/30G01N1/34G01N33/533G01N2001/305
Inventor 王淑娟刘文举刘晓丽庞训胜闻爱友华金玲
Owner ANHUI SCI & TECH UNIV
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