Mixed fluorescent dye reagent and freeze-drying method thereof

Abstract

The invention provides a mixed fluorescent dye reagent. Themixed fluorescent dye reagent comprises, by volume, 71-96 ml of dimethyl sulfoxide and 3-29 ml of SYTOX Red, and further comprises, by mass, 0.03-0.65 g of DCFH-DA and 0.01-0.1 g of MitoSOX Red. The invention further provides a freeze-drying method of the mixed fluorescent dye reagent. A freeze-dried product of the mixed fluorescent dye reagent obtained by the method has the characteristics of complete and regular appearance, no collapse, 14 days of storage time, 20 h of storage time after redissolution, and small performance loss, the quality of cells including germ cells after redissolution can be detected, such as the cell active oxygen damage ratio and the cell death ratio, and the O2<-> and H2O2 active oxygen damage can be specifically detected;.

Description

technical field [0001] The invention relates to the field of fluorescent dyes, in particular to a mixed fluorescent dye reagent and a freeze-drying method thereof. Background technique [0002] Active oxygen in the body, such as O 2 - and H 2 o 2 It has the characteristics of high reactivity, so they will cause oxygen damage to sperm. In addition, the life of active oxygen is short, and the requirements for detection reagents and detection methods are high. At the same time, in the detection of O 2 - and H 2 o 2 When reactive oxygen species are damaged, other types of reactive oxygen species will affect the detection accuracy of the above-mentioned reactive oxygen species because there is no specific selection of detection reagents currently provided. [0003] In addition, there are still dead sperm. Therefore, the existence of sperm is more. The current dye reagents and testing methods cannot exclude the interference of other active oxygen, and can also detect the su...

AI Technical Summary

Problems solved by technology

[0002] Active oxygen in the body, such as O 2 - and H 2 o 2 It has the characteristics of high reactivity, so they will cause oxygen damage to sperm. In addition, the life of active oxygen is short, and the requirements for detection reagents and detection methods are high. At the same time, in the detection of O 2 - and H 2 o 2 When active oxygen is damaged, other types of active oxygen will affect the detection accuracy of the above active oxygen because there is no specific selection of detection reagents currently provided

[0003] In addition, there are still dead sperm. Therefore, the existence of sperm is relatively large. The current dye reagents and testing methods cannot exclude the interference of other reactive oxygen species, and can also detect the survival rate of sperm

[0004] In addition, fluorescent dye reagents usually have poor performance stability. Therefore, in order to maintain stable performance of fluorescent dyes for a long time, the fluorescent dyes need to be lyophilized. The components and ratios in the mixed fluorescent dye reagents will affect the freeze-dried shape of the mixed fluorescent dye reagents. Performance, storage limit, and also affect the performance and storage time after reconstitution

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