3D dynamic observation of human sperm by parallel phase-shifting digital holographic microscopy based on pixelated polarization

Morphology and motility are essential criteria for assessing sperm viability. However, the human head is small (∼3–4 μm) requires a relatively high-magnification microscope objective, while flagella (∼45 poorly visible with complex 3D properties. Microscopic dynamic observation of intact in challenging. Conventional inspection methods limited depth field inadequate this issue. To provide solution to critical need, we develop pixelated polarization-based parallel phase-shifting digital holographic microscopy sperm. Compared conventional imaging, approach can effectively separate object wavefront avoid image quality degradation fully exploiting spatial bandwidth camera. We propose use Stokes parameter reconstruction method reconstruct investigate effect sampling interval on system resolution by spectral analysis. The methodology achieves retrieval trajectory motion parameters reconstructs orientation thin, highly-dynamic flagellum. allows more comprehensive information morphology, which significant male reproductive research. It also has potential micro-organisms.