A Simple and Rapid Biochemical Method for Measuring ROS in Infertile Semen Sample- A Technical Report

Determining reactive oxygen species (ROS) can be challenging when the study subject is infertile, as ROS free radicals are essential to sperm quality. We offer a simple approach that employs a basic sperm detergent mixture (SDM) to assess semen ROS levels both colourimetrically and qualitatively (i.e., the color of the sperm cell exposed to ROS) using microscopy. Reactive oxygen species levels in samples from both fertile and infertile semen are assessed using SDM, which comprises the following procedures: a) Get ready: Separate sperm cells for qualitative assessment and prepare viable and infertile samples by WHO criteria. Lyse sperm cells in lysis buffer, then centrifuge the pellets at 500 g. Pellets were washed first with NBT solution, then with a phosphate-buffered solution; d) the reaction was terminated by adding 2M KOH and DMSO; and e) the final color development was measured at 590 nm. The obtained results show that early and ideal to determine ROS in infertile semen instead of other kits/staining procedures. It is also less expensive than the chemiluminescence test and ideal for use in a small-scale andrology or teaching laboratories.