Improved crystal violet staining method for detecting sperm acrosome

A technology of crystal violet and dyeing method, which is applied in the preparation of test samples, measuring devices, sampling, etc., can solve the problems of complicated and time-consuming operation, expensive reagents, etc., and achieve the effects of wide sources, convenient preparation and easy operation.

Inactive Publication Date: 2018-03-16
高晓勤 +1
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Problems solved by technology

[0003] To sum up, the problems existing in the existing technologies are: most of them cannot be po

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  • Improved crystal violet staining method for detecting sperm acrosome
  • Improved crystal violet staining method for detecting sperm acrosome
  • Improved crystal violet staining method for detecting sperm acrosome

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[0015] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0016] The application principle of the present invention will be described in detail below in conjunction with the accompanying drawings.

[0017] Such as figure 1 As shown, the improved crystal violet staining method for detecting sperm acrosomes provided by the embodiments of the present invention includes the following steps:

[0018] S101: Use 20ml of 5% gelatin aqueous solution, 50ml of 0.05% glutaraldehyde solution, 100ml of pH7.0 barbian acetate buffer solution; 50ml of 0.5% trypan blue solution; 100ml of pH7.3 phosphate buffer solution; 500ml of double distilled water Preparation of modified gelatin base film;

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Abstract

The invention belongs to the technical field of sperm acrosome detection, and discloses an improved crystal violet staining method for detecting sperm acrosome. The method comprises the following steps: preparing an improved gelatin film; liquefying collected fresh seminal fluid, subjecting seminal fluid to centrifugation, discarding seminal plasma, suspending precipitates in a phosphate buffer solution, placing the buffer solution in a thermotank, evenly paving sperm suspension on the gelatin film, incubating the sperms in a thermotank with a temperature of 37 DEG C, after 30 minutes, takingout sperm suspension for microscopic examination; dropwise adding a crystal violet dyeing solution on a fixed smear, carrying out dyeing for 1 to 3 minutes, washing the smear by water to remove excessdyes; dropwise adding a Lugol iodine solution, after one minute, washing the smear by water, then dropwise adding alcohol (95%) to perform color separation, shaking the glass slide until the purple color is not faded, washing the smear by flowing water, and carrying out microscopic examination after drying. A simple and rapid dyeing medium adopted by the crystal violet staining method can enhancethe cell dyeing affinity, through a discoloring treatment, the binding strength between a fuel and a polluted object can be measured, the structural difference of cells can be discriminated, and thesituation of sperm acrosome is well displayed.

Description

technical field [0001] The invention belongs to the technical field of sperm acrosome detection, and in particular relates to a method for detecting sperm acrosome by an improved crystal violet staining method. Background technique [0002] The sperm acrosome is formed by the Golgi complex near the sperm nucleus, and the acrosome is attached to the front surface of the nucleus in a vesicular hemispherical shape. Sperm acrosomes are rich in polysaccharides and various hydrolytic enzymes, which are significantly different from those contained in the nucleus. The improved simple, rapid and mordant crystal violet staining method can enhance the staining affinity of cells, and then decolorize to measure the binding fastness between the dye and the dyed object. In order to identify the difference between the cell structure, better show the presence of sperm and acrosome. With the development of science, biochemical examination techniques have good results in the determination of...

Claims

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Application Information

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IPC IPC(8): G01N1/30
CPCG01N1/30G01N2001/302
Inventor 高晓勤戴研平杨薇杨喆马晓萍李淑钗岳应权王平
Owner 高晓勤
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