The left testis of mice was fixed with formaldehyde fixative, embedded, sliced, and stained with hematoxylin and eosin (H&E), and the images were collected. The left and right epididymis of the mice were cut into pieces in an EP tube containing normal saline, and then shaken and incubated in a water bath at 37 °C for 15 min [32 (link)]. The sperm quality was analyzed according to the method of Oghbaei et al. [33 (link)]. A drop of diluted diluent was taken on the blood cell count, and the number of sperm in five squares was counted under a biological microscope (CX33, Olympus Corporation, Tokyo, Japan). The number of sperm per milliliter was equal to N × dilution factor × 5 × 104. Another drop of diluent was taken to smear; 200 sperms were observed, and the number of active sperm was counted. In addition, the sperm was stained according to the instructions of the quick sperm stain kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China), and a fluorescence-inverted microscope–spectrometer coupling (DMI4000B+Iso Plane 160, Leica Corporation, Wetzlar, Germany) was used to observe and capture the stained sperm images.
Quick sperm stain kit
Manufactured by Nanjing Jiancheng
Sourced in China
The Quick Sperm Stain Kit is a laboratory product designed for the rapid staining of sperm cells. It provides a simple and efficient method for the visualization and identification of sperm under a microscope. The kit includes the necessary reagents and instructions for performing the staining procedure.
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Lab products found in correlation
5 protocols using quick sperm stain kit
1
Sperm Quality Analysis in Mice
2
Epididymal Sperm Staining Procedure
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The sperm was collected from cauda epididymides and placed in physiological saline (37 °C). The sperm was stained by Quick sperm stain kit (Nanjing Jiancheng Bioengineering Institute Co., Ltd., Nanjing, China) and eosin stain kits (Zhuhai Beisuo Biotechnology Co. LTD., Zhuhai, China). Finally, the staining of sperm was observed with an EVOS imaging system (Invitrogen, USA).
3
Sperm Morphology Evaluation by Staining
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The sperm was collected from cauda epididymides and placed in physiological saline (37 °C). The sperm was stained by Quick sperm stain kit (Nanjing Jiancheng Bioengineering Institute Co., ltd., Nanjing, China). The head, mid-piece and tail of sperm were observed with an EVOS imaging system (Invitrogen, USA).
4
Epididymal Sperm Analysis Protocol
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The entire right epididymis was placed in 1 mL of preheated saline and cut into small segments. After incubation in a 37 °C environment, the sperm suspension was dropped into a Neobar’s hemocytometer and the sperm count was estimated under a coverslip. The rate of abnormal sperm morphology was assessed and calculated by observing the sperm staining images using the Quick sperm stain kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). Sperm parameters were analyzed according to the method adopted by Qiu et al. [29 (link)].
5
Spermatozoa Evaluation Protocol
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The entire right epididymis was placed in 1 mL of prewarmed saline and minced into small sections, incubating for 10 min (37 °C, 5% CO2) to allow the spermatozoa to swim out from the epididymal tubules. The sperm suspension was dropped into a Neobar’s hemocytometer, and the sperm count was estimated under a cover glass. The sperm motility was observed under a 400× light microscope (Olympus Corporation, Tokyo, Japan) and calculated and expressed as a percentage of motile sperm according to the World Health Organization manual criteria. The sperms were stained using the Quick sperm stain kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to the manufacturer’s instructions. Additionally, images of sperm after staining were observed under a light microscope and acquired by photographing using a Leica DMI4000B microscope with Leica DFd4500 imaging system (Leica Corporation, German). The rate of abnormal morphology of sperm was evaluated and calculated by observing sperm staining images. Sperm parameters of mice (count, motility, and morphology) were analyzed in each group according to the methods adopted by Qiu et al. and Oghbaei et al. [49 (link),50 (link)].
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