Bioluminescence detection of intracellular ATP in porcine sperm cells using paper-based devices

Koji Matsuura, Zheyi Chen, Shuyu Hu, Yuka Asano, Chao-Min Cheng

Mammalian sperm motion relies on adenosine triphosphate (ATP) to produce energy for dynein slide-motion via hydrolysis. To clarify the relationship between mammalian sperm motility and ATP production/consumption, semen ATP concentration was examined using a paper-based bioluminescence assay (luciferase) due to its higher sensitivity, reduced detection time, and minimal reagent requirements compared to fluorometry or colorimetry. ATP concentration in the form of sperm sample signal intensity was then correlated to motile sperm count and sperm motility, with higher ATP concentrations indicating the presence of lower-motility sperm and lower ATP concentrations indicating higher ATP consumption and higher-motility sperm. Employing the described paper-based bioluminescence methodology and selective inhibition of glycolysis and oxidative phosphorylation (OXPHOS), porcine sperm was found to produce more ATP through OXPHOS than glycolysis. The results of our analyses support the notion that bioluminescence-based ATP detection on a paper-based device can be employed to examine the contribution of these two metabolic systems to sperm motility and fertilization.

Microfluidics and Nanofluidics

Springer Nature

30

42

https://doi.org/10.1007/s10404-026-02894-z