Impact of Micro and Nano Zinc Oxide Particles on Lentil Seed’s Internal Activity Using Biospeckle Optical Coherence Tomography (bOCT)

The increasing use of nanoparticles (NPs) in various industries has intensified research into plant–NP interactions. NP properties significantly impact their cellular uptake and plant effects, highlighting the need for advanced monitoring techniques to understand their influence on plant growth and seed germination. This study uses biospeckle optical coherence tomography (bOCT) to investigate the size-dependent effects of zinc oxide (ZnO) NPs and microparticles (MPs) on lentil seed internal activity, visualizing dynamic changes under ZnO particle stress. ZnO was selected for its agricultural relevance as a micronutrient. Lentil seeds were submerged in ZnO particle dispersions (<50 nm, <100 nm, 5 μm, 45 μm) at concentrations of 0 (control), 25, 50, 100, and 200 mg/L. OCT structural images were obtained at 12.5 frames per second using a swept-source OCT (central wavelength 1.3 μm, bandwidth 125 nm, sweep frequency 20 kHz). OCT scans were performed before immersion (0 h) and 5, 10, and 20 h after lentil seed exposure to particle dispersion. The biospeckle image, representing dynamic speckle patterns characteristic of biological tissues, was calculated as the ratio of standard deviation to mean of 100 OCT structural images over 8 s. Biospeckle contrast was compared 0, 5, 10, and 20 h post-exposure. ZnO NPs <50 nm and 100 nm negatively impacted lentil seed biospeckle contrast at all concentrations. In contrast, 45 µm ZnO MPs significantly increased it even at 100 mg/L, while 5 μm MPs decreased biospeckle contrast at higher concentrations. bOCT results were compared with conventional morphological (germination percentage, growth, biomass) and biochemical (superoxide dismutase, catalase, and hydrogen peroxide) measurements. Conventional methods require one week, whereas bOCT detects significant changes in only five hours. The results from bOCT were consistent with conventional measurements. Unlike standard OCT, which monitors only structural images, bOCT is capable of monitoring internal structural changes, allowing rapid, non-invasive assessment of nanomaterial effects on plants.