Pretreatment method for keeping animal sperm motility and determining animal sperm apoptosis
A treatment method and technology of sperm motility, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problems of long pretreatment methods, affecting sperm apoptosis analysis, and easy death.
Active Publication Date: 2022-05-10
SHANGHAI INNOSTAR BIO TECH
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Problems solved by technology
[0004] In order to overcome the defects in the prior art that the sperm are easy to die in the pretreatment process of the flow cytometer analysis after being separated from the body, which affects the analysis of sperm apoptosis, and the pretreatment method of the prior art takes a long time, a method is provided. Treatment method for maintaining animal sperm vitality and a pretreatment method for measuring animal sperm apoptosis applied to flow cytometry
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Embodiment 1
[0055] The incubation solution of Example 1: 0.01% bovine serum albumin solution, 100mmol / L HEPES / NaOH, containing 1.4mmol / L NaCl and 25mmol / L CaCl2, pH7.5.
Embodiment 2
[0056] The incubation solution of Example 2: 0.00125% bovine serum albumin solution, 100mmol / L HEPES / NaOH, containing 1.4mmol / L NaCl and 25mmol / L CaCl2, pH7.5.
Embodiment 3
[0057] The incubation solution of Example 3: 0.005% bovine serum albumin solution, 100mmol / L HEPES / NaOH, containing 1.4mmol / L NaCl and 25mmol / L CaCl2, pH7.5.
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The invention discloses a treatment method for keeping animal sperm motility and a pretreatment method for animal sperm apoptosis determination. The treatment method for keeping animal sperm motility comprises the following steps: incubating cut animal epididymis in an incubation solution; the incubation liquid is a combined liquid of 0.00125-0.01% of bovine serum albumin liquid and an HEPES / NaOH buffer solution, and the pH value of the incubation liquid is 7.2-7.7. The pretreatment method for determining the apoptosis of the animal sperms comprises the following steps: filtering the incubated suspension, and taking sperm filtrate; and dyeing the sperm filtrate. According to the treatment method for keeping the activity of the animal sperms, the isolated sperms are incubated in a proper environment, the incubation liquid is optimized, the incubation time is shortened, and the sperms keep the activity. The pretreatment method for determining the apoptosis of the animal sperms applied to the flow cytometry also comprises the step of effectively shortening the flow pretreatment time without influencing the subsequent fluorescence labeling and analysis effects.
Description
technical field [0001] The invention relates to a treatment method for maintaining the vitality of animal sperm and a pretreatment method for measuring the apoptosis of animal sperm applied to a flow cytometer. Background technique [0002] In male reproductive toxicology research, sperm motility is an important observation index, and flow cytometry is a multi-parameter, rapid quantitative analysis of single cells or other biological particles at the molecular level. It can analyze tens of thousands of cells at high speed, and can measure multiple parameters from one cell at the same time. It has the advantages of high speed, high precision and good accuracy. It is one of the most advanced cell quantitative analysis technologies in the contemporary era. The use of flow cytometry to detect animal sperm motility is a relatively sensitive detection method, which can detect early apoptosis, mechanical death, and late apoptosis of sperm. However, in the pretreatment process of fl...
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IPC IPC(8): G01N1/28G01N1/30G01N1/34G01N15/14C12N5/076
CPCG01N1/28G01N1/30G01N1/34G01N15/14C12N5/061C12N2501/998C12N2500/60Y02A40/81
Inventor 常艳洪敏李英奇李嫚琪杨紫轩许瀚林王欣
Owner SHANGHAI INNOSTAR BIO TECH
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