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Fluorescent staining method of evaluating sperm motility of tree shrew

A technology of sperm motility and fluorescent dyeing, applied in the field of animal reproductive biology, can solve the problems of undiscovered and less data, and achieve the effect of accurate detection

Inactive Publication Date: 2015-04-08
YUNNAN AGRICULTURAL UNIVERSITY
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Problems solved by technology

Now, more and more people of insight believe that the tree shrew is the most promising new type of experimental animal to replace the precious primates, and the research on the reproductive biology of the tree shrew will help the artificial reproduction of this kind of animal. So far, there are very few data about tree shrew reproductive biology, and no report identical or similar to the present invention has been found so far

Method used

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Embodiment

[0021] Each 100mL solution contains 742 mg sodium chloride, 23.6 mg potassium chloride, 29.4 mg calcium chloride dihydrate, 10.2 mg magnesium chloride hexahydrate, 90.1 mg glucose, 4.8 mg sodium dihydrogen phosphate, 16.8 mg sodium bicarbonate, acetone The ratio of 1.1 mg sodium lactate, 1 mg phenol red, 130.2 mg HEPES sodium, 119.2 mg HEPES acid, 185 μL 60% aqueous solution of sodium lactate, 5 mg gentamicin, and 300 mg bovine serum albumin was obtained by using Milli-Q ultrapure Mix the water evenly, adjust the pH to 7.4 with NaOH or HCl, make TALP-HEPES semen washing solution, and store it in a water bath at 37 °C; separate the cauda epididymis and vas deferens from the tree shrew testis by surgical operation, wash the epididymis with TALP-HEPES and remove Remove blood and adipose tissue, then place the cauda epididymis in a small centrifuge tube containing 1 mL of 37 ℃ TALP-HEPES, cut the cauda epididymis with fine surgical scissors, and continue to incubate at 37 ℃ for ...

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Abstract

The invention provides a fluorescent staining method of evaluating sperm motility of a tree shrew. The fluorescent staining method comprises the following steps: (1) preparing a TALP-HEPES sperm rinse; (2) collecting sperms; (3) washing the sperms; (4) staining; and (5) detecting by a microscope. According to the fluorescent staining method provided by the invention, by utilization of the characteristics that Hoeches 333422 and PI are different in reaction to dead and alive sperms and can be excited by a laser with a same wavelength, fluorescent staining is carried out on sperms of the tree shrew, and the sperm motility can be accurately detected without needing to switch a light source; in addition, as the two dyes respectively emit red and blue fluorescence after being excited by the laser, a dye which emits green fluorescence can also be introduced to detect the integrity of sperm acrosomes while the motility is detected, and the sperm motility and the integrity of the sperm acrosomes of the tree shrew are accurately detected by virtue of a fluorescence microscope or a flow cytometer.

Description

technical field [0001] The invention belongs to the technical field of animal reproductive biology, in particular to a fluorescent staining method for evaluating tree shrew sperm vitality. Background technique [0002] Tree shrews belong to the order of climbing shrews, which is a class of small mammals similar in shape to squirrels. Although humans have studied tree shrews for 200 years, it was not until the 1980s that tree shrews were used as model animals in biology. Research has attracted more attention. Tree shrews are highly similar to primates and even humans in terms of physiological anatomy, neurodevelopment, and psychological stress, and they also have small size, short reproductive cycle, fast reproduction, and low feeding costs. It is widely used in many fields of biology and medical research. Now, more and more people of insight believe that the tree shrew is the most promising new type of experimental animal to replace the precious primates, and the resea...

Claims

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Application Information

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IPC IPC(8): C12Q1/06
Inventor 李亚辉杨明华
Owner YUNNAN AGRICULTURAL UNIVERSITY
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