Related Experiment Video
Updated: Aug 16, 2026
A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
Published on: April 11, 2014
Ascorbic acid mitigates oxidative structural degradation in bovine spermatozoa: a label-free quantitative phase
Harpreet Kaur1,2, Shubham Tiwari1,2, Aman Kumar3
1Center for Sensors, Instrumentation and Cyber Physical System Engineering (SeNSE), Indian Institute of Technology Delhi, Hauz Khas, New Delhi, 110016, India.
None:
Oxidative stress is a key factor in low fertility outcomes during assisted reproduction technology (ART) and contributes to poor sperm quality. Conventional sperm assessment relies on bright-field microscopy, which lacks the quantitative sensitivity to resolve subcellular structural and biophysical changes without exogenous contrast agents, a requirement that can introduce cytotoxic effects and compromise cell viability. This study uses quantitative phase microscopy (QPM) as a label-free, high-throughput method to demonstrate how ascorbic acid (Vitamin C) reduces the impact of oxidative structural degradation in Sahiwal bovine spermatozoa. To illustrate this pathology, severe oxidative stress was experimentally induced using hydrogen peroxide (H2O2), and two doses of ascorbic acid (1 mg/ml and 8 mg/ml) were tested to evaluate dose-dependent antioxidant recovery in structurally damaged sperm. QPM was used to quantify structural changes in key biophysical parameters, including dry mass, optical thickness, volume, surface area, sphericity, and surface area-to-volume ratio, as well as intracellular texture parameters. Under acute oxidative stress, bovine spermatozoa exhibited significant reductions in optical thickness and dry mass, alongside measurable changes in intracellular structural organization, collectively indicative of oxidative stress-induced morphological degradation. Co-treatment with ascorbic acid resulted in partial, dose-dependent structural preservation, with the 8 mg/ml formulation demonstrating statistically significant attenuation of these structural changes compared to the 1 mg/ml treatment group. These findings suggest that QPM-derived biophysical parameters may serve as promising, label-free structural indicators for characterizing oxidative damage in bovine spermatozoa. Future studies should incorporate functional validation to determine whether this structural preservation translates to improved outcomes in ART.
