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We report a multilayer lensless in-line holographic microscope (LIHM) with improved imaging resolution by using the pixel super-resolution technique and random sample movement. In our imaging system, a laser beam illuminated the sample and a CMOS imaging sensor located behind the sample recorded the in-line hologram for image reconstruction. During the imaging process, the sample was moved by hand randomly and the in-line holograms were acquired sequentially. Then the sample image was reconstructed from an enhanced-resolution hologram obtained from multiple low-resolution in-line holograms by applying the pixel super-resolution (PSR) technique. We studied the resolution enhancement effects by using the U.S. Air Force (USAF) target as the sample in numerical simulation and experiment. We also showed that multilayer pixel super-resolution images can be obtained by imaging a triple-layer sample made with the filamentous algae on the middle layer and microspheres with diameter of 2 μm on the top and bottom layers. Our pixel super-resolution LIHM provides a compact and low-cost solution for microscopic imaging and is promising for many biomedical applications.

Multilayer pixel super-resolution lensless in-line holographic microscope with random sample movement