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A mouse embryo blastocyst staining method for quality control of human assisted reproductive consumable products

A technology for assisted reproduction and mouse embryos, which is applied in the preparation of test samples, measuring devices, and analysis materials, etc., can solve the problems of high cost of quality control, complicated technical procedures, etc. Solve the effect of high cost of quality control

Inactive Publication Date: 2020-06-12
东蕴医疗科技(上海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method uses bis-benzimide (Hoechst33342) reagent, DABCO (1,4-Diazabicyclo[2.2.2]octane solution, DABCO) anti-fade reagent, anti-UTF1 antibody, and the corresponding bismuth of anti-UTF1 antibody Advanced reagents such as anti-antibody and DNA staining solution are required, and a stereo microscope and a fluorescence microscope are required to be used together; this method is not only complicated in technical procedures, but also has disadvantages such as high cost of quality control

Method used

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  • A mouse embryo blastocyst staining method for quality control of human assisted reproductive consumable products
  • A mouse embryo blastocyst staining method for quality control of human assisted reproductive consumable products

Examples

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Embodiment 1

[0021] Example 1: A mouse embryo blastocyst staining method for quality control of human assisted reproduction consumable products, the staining operation is as follows:

[0022] ⑴Fixation: Take 3-5 blastocysts and place them in the center of the glass slide to form droplets with a diameter of 3-5mm (20-30ul), wash with phosphate buffer solution with a pH value of 7.0-7.5 for 1 minute, and then change to 4.0 % paraformaldehyde (20-30ul) was fixed for 5 minutes, 4.0% paraformaldehyde was sucked off, and 4.0% paraformaldehyde (20-30ul) was added dropwise to fix for 5 minutes. Wash with purified water (20-30ul) with a pH value of 5.0-7.0 for 1 minute, absorb excess liquid, wash with water three times, and proceed to the next step.

[0023] (2) Staining: add 20ul of hematoxylin staining solution, observe under a microscope, and control the staining time to 3 minutes according to the staining of blastocyst cell nuclei. Wash with purified water with a pH value of 5.0 to 7.0 for 1 m...

Embodiment 2

[0027] Example 2: A mouse embryo blastocyst staining method for quality control of human assisted reproduction consumable products, the staining operation is as follows:

[0028] ⑴Fixation: Take 3-5 blastocysts, place them in the center of the glass slide, form a droplet with a diameter of 3-5mm (20-30ul), wash with phosphate buffer solution with a pH value of 7.0-7.5 for 3 minutes, and then change to 5.0 % paraformaldehyde (20-30ul) was fixed for 10 minutes, and 5.0% paraformaldehyde was absorbed. Wash with purified water (20-30 ul) with a pH value of 5.0-7.0 for 3 minutes, absorb excess liquid, wash twice with water repeatedly, and proceed to the next step.

[0029] (2) Staining: add 20ul of hematoxylin staining solution, observe under a microscope, and control the staining time to 5 minutes according to the staining of blastocyst cell nuclei. Wash with purified water with a pH value of 5.0 to 7.0 for 3 minutes, absorb excess liquid, wash twice with water repeatedly, and pl...

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Abstract

The invention provides a mouse embryo blastocyst staining method for quality control of human assisted reproductive consumable products. The method comprises the following steps of culturing a mouse embryo in vitro until a blastocyst is formed; observing the number of blastocyst cells through mouse embryo blastocyst staining operation, wherein the staining operation comprises the following steps:fixing: flushing the mouse blastocyst cells with a buffer solution with a specific pH value before staining, fixing with paraformaldehyde with the concentration of 3.0-5.0% for 5-10 minutes, and flushing with a buffer solution with a specific pH value; dyeing: dyeing the mouse blastocyst with hematoxylin for 2-5 minutes after the mouse blastocyst is fixed, and then washing with purified water; color separation and bluing: after dyeing the mouse blastocyst, carrying out color separation with 0.5-1.5% hydrochloric acid alcohol for 5-30 seconds, and then washing with purified water; after washing, bluing for 5-15 minutes by using 1.0-3.0% ammonia water, and then washing by using purified water; fixing and sealing: after the mouse blastocyst is blued, dropwise adding anhydrous glycerol, covering with cover glass, adding nail polish for sealing, and the like. Dyeing result judgment: blastocyst cells are uniformly dispersed, and dyeing is clear.

Description

technical field [0001] The invention relates to a quality control technology for human assisted reproduction consumable products, in particular to a method for staining mouse embryo blastocysts. Background technique [0002] Human in vitro assisted reproductive technology (Assisted Reproductive Technology, ART) refers to taking out eggs and sperm, placing them in vitro for fertilization (IVF, in vitro fertilization) or through intracytoplasmic sperm injection (ICSI), using artificial The method is to fertilize eggs and sperm in vitro and carry out early embryo development, and then transplant them into the mother's uterus to develop and give birth to babies. It is an important technical means to solve infertility and even achieve eugenics. In the process of ART, a series of medical devices are needed to realize the process of gamete acquisition, fertilization, development, etc., so that they can be safely applied to ART. It is required to use the blastocyst counting method ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30G01N1/28
CPCG01N1/30G01N1/28G01N2001/302
Inventor 胡彦新尹航夏旭升于跃龙
Owner 东蕴医疗科技(上海)有限公司