Better understanding of particle–particle and particle–fluid interactions requires accurate 3D measurements of particle distributions and motions. We introduce the application of in-line digital holographic microscopy as a viable tool for measuring distributions of dense micrometer \left(3.2\text{\hspace{0.17em} μm}\right) and submicrometer \left(0.75\text{\hspace{0.17em} μm}\right) particles in a liquid solution with large depths of 1–10\text{\hspace{0.17em} mm} . By recording a magnified hologram, we obtain a depth of field of \sim 1000 times the object diameter and a reduced depth of focus of approximately 10 particle diameters, both representing substantial improvements compared to a conventional microscope and in-line holography. Quantitative information on depth of field, depth of focus, and axial resolution is provided. We demonstrate that digital holographic microscopy can resolve the locations of several thousand particles and can measure their motions and trajectories using cinematographic holography. A sample trajectory and detailed morphological information of a free-swimming copepod nauplius are presented.

© 2006 Optical Society of America

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