Sperm Count and Motility Quantification

The total sperm count and percentage of sperm motility were determined as described previously [20 (link)]. In brief, the distal cauda epididymis was excised and transferred to a 1.5 mL microcentrifuge tube containing normal saline, nicked at two sites with scissors. Spermatozoa were allowed to suspend in pre-warmed normal saline for 5 min at 37 °C. The total sperm count was calculated by counting the number of sperm in 10 squares of the grid after adding two drops of each specimen onto a counting chamber (SEFI-Medical Instruments Ltd., Hicksville, NY, USA) under an Axio Imager 2 light microscope (Carl Zeiss MicroImaging LLC, Goettingen, Germany) at 20× magnification. Sperm count is expressed as 10⁶ sperms/mL. Sperm motility was evaluated under a light microscope at 20× magnification within 10 squares of the grid on a pre-warmed counting chamber. The percentage of motile spermatozoa was analyzed by the formula ((mean number of motile spermatozoa/total number of spermatozoa) × 100) and the results are given as a percentage.

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Karna K.K., Soni K.K., You J.H., Choi N.Y., Kim H.K., Kim C.Y., Lee S.W., Shin Y.S, & Park J.K. (2020). MOTILIPERM Ameliorates Immobilization Stress-Induced Testicular Dysfunction via Inhibition of Oxidative Stress and Modulation of the Nrf2/HO-1 Pathway in SD Rats. International Journal of Molecular Sciences, 21(13), 4750.

Publication 2020

Axio Imager 2 light microscope

Epididymis Microscope light Normal saline Sperm motility Spermatozoa

Corresponding Organization : Jeonbuk National University

Other organizations : Kent State University, Kyungsung University, Hanyang University, Samsung Medical Center, Sungkyunkwan University

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Variable analysis

independent variables

Cauda epididymis excision and transfer to microcentrifuge tube

dependent variables

Total sperm count
Percentage of sperm motility

control variables

Normal saline used to suspend spermatozoa
Incubation temperature of 37 °C
Counting chamber (SEFI-Medical Instruments Ltd., Hicksville, NY, USA)
Axio Imager 2 light microscope (Carl Zeiss MicroImaging LLC, Goettingen, Germany) at 20× magnification

controls

No positive or negative controls explicitly mentioned

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