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Flow cytometry-based sperm acrosome reaction detection reagent

A technology of flow cytometry and acrosome reaction, which is applied in the field of sperm acrosome reaction detection reagents based on flow cytometry, can solve the problems of cumbersome operation process, difficult promotion and application, and large sperm damage, and achieve accurate detection results, Capabilization effect is good, easy to induce the effect

Inactive Publication Date: 2013-10-02
ZHEJIANG CELLPRO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the detection results of such methods are too dependent on human operation, and the detection results are highly subjective; the operation process is cumbersome, the detection efficiency is low, and the repeatability is poor; The requirements for semen samples are too high, it is not suitable for routine testing, and it is difficult to promote clinical application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] Embodiment 1, a kind of sperm acrosome reaction detection reagent based on flow cytometry, is made up of four parts of acrosome reaction inducing liquid, sperm capacitation liquid, FITC-PSA fluorescent staining liquid and propidium iodide staining liquid, wherein The acrosome reaction induction solution consists of 5mg Ca 2+ Vehicle A23187 was obtained by dissolving in 9.56 mL DMSO. Sperm capacitation fluid consists of sodium chloride, potassium dihydrogen phosphate, glucose, potassium chloride, magnesium sulfate heptahydrate, Hepes-Na, sodium bicarbonate, sodium pyruvate and BSA in a reagent bottle, add distilled water to fully dissolve and mix, Then add calcium chloride dihydrate solution and sodium lactate solution with a concentration of 600g / L, adjust the pH value to 7.4 with 1M HCl, and then filter with a 22μm filter membrane to form a solution with a concentration of sodium chloride of 0.12mol / L and a concentration of potassium dihydrogen phosphate The concentra...

Embodiment 2

[0010] Embodiment 2 is basically the same as Embodiment 1, except that in the sperm capacitation fluid, the concentration of sodium chloride is 0.05 mol / L, the concentration of potassium dihydrogen phosphate is 0.5 mmol / L, the concentration of glucose is 2.0 mmol / L, The concentration of potassium chloride is 2.3 mmol / L, the concentration of magnesium sulfate heptahydrate is 0.2 mmol / L, the concentration of calcium chloride dihydrate is 0.8 mmol / L, the concentration of Hepes-Na is 0.01 mol / L, and the concentration of sodium bicarbonate is 2.4 mmol / L, the concentration of sodium pyruvate is 0.1 mmol / L, the mass percent concentration of BSA is 2%, and the volume of sodium lactate solution is 0.15%. In propidium iodide staining solution, the concentration of sodium chloride is 50mmol / L, the concentration of potassium chloride is 2.3 mmol / L, the concentration of disodium hydrogen phosphate is 3.2mmol / L, the concentration of potassium dihydrogen phosphate is 0.5mmol / L, PI The conce...

Embodiment 3

[0011] Embodiment 3 is basically the same as Embodiment 1, except that in the sperm capacitation fluid, the concentration of sodium chloride is 0.19 mol / L, the concentration of potassium dihydrogen phosphate is 2.0 mmol / L, and the concentration of glucose is 10.5 mmol / L. The concentration of potassium chloride is 7.8 mmol / L, the concentration of magnesium sulfate heptahydrate is 2.4 mmol / L, the concentration of calcium chloride dihydrate is 3.4 mmol / L, the concentration of Hepes-Na is 0.04 mol / L, and the concentration of sodium bicarbonate is 5.6 mmol / L, the concentration of sodium pyruvate is 0.5 mmol / L, the mass percent concentration of BSA is 10%, and the volume of sodium lactate solution is 0.75%. In propidium iodide staining solution, the concentration of sodium chloride is 190mmol / L, the concentration of potassium chloride is 7.8 mmol / L, the concentration of disodium hydrogen phosphate is 10mmol / L, the concentration of potassium dihydrogen phosphate is 2.0mmol / L, and the...

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Abstract

The invention discloses a flow cytometry-based sperm acrosome reaction detection reagent which comprises acrosome reaction induction fluid and staining fluid, as well as sperm capacitation fluid, the pH value of the sperm capacitation fluid is 7.0-7.8, the sperm capacitation fluid contains sodium chloride, potassium dihydrogen phosphate, glucose, potassium chloride, magnesium sulfate heptahydrate, calcium chloride dehydrate, N-2-piperazine-N-2-ethanesulfonic acid disodium salt, sodium bicarbonate, sodium acetone, bovine serum albumin, and sodium lactate solution, the sperm capacitation fluid replaces the current sperm culture fluid, so that the capacitation effect is good and the induction is facilitated; the sperm acrosome reaction can be detected by using flow cytometry, and the flow cytometry-based sperm acrosome reaction detection reagent is accurate in the sperm acrosome reaction detection result, stable and reliable, good in repeatability, and clinically easily popularized.

Description

technical field [0001] The invention relates to a sperm acrosome reaction detection reagent, in particular to a sperm acrosome reaction detection reagent based on flow cytometry. Background technique [0002] Semen analysis is an indispensable part of infertility examination, mainly including sperm concentration, motility and morphology examination, etc. However, these examination indicators cannot fully evaluate male fertility; and some male infertility patients are not found in routine semen examination Normal, therefore, it is necessary to introduce new indicators for sperm function detection and evaluation. The acrosome is a structure unique to sperm. During the process of recognizing and combining with the zona pellucida of the egg, the acrosome is induced to undergo an acrosome reaction, thereby realizing the fusion of sperm DNA and egg DNA. Therefore, the fertilization ability of sperm can be evaluated by detecting the ability of sperm to undergo acrosome reaction. ...

Claims

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Application Information

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IPC IPC(8): G01N15/14
Inventor 江南曾桥薛志刚
Owner ZHEJIANG CELLPRO BIOTECH
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