The flagellum is vital for eukaryotic cell survival. We propose method utilizing digital holographic microscopy (DHM) advantages, such as high-speed imaging and precise axial localization, to develop a versatile approach for four-dimensional tracking (X, Y, Z, time) of eukaryotic microorganisms' bodies and flagella, addressing challenges faced by existing methods. We reconstructed for the first time the shape of a 200 nm diameter Chrysochromulina simplex flagellum and measured mouse sperm flagella over time, capturing approximately 800 points. Our technique opens new avenues for studying flagella's roles in cellular functions and survival strategies, offering high-speed and precise 3D tracking at the nanoscale.
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