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

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.

Resolution enhancement method for lensless in-line holographic microscope with spatially-extended light source.

We propose a resolution enhancement method for lensless in-line holographic microscope (LIHM) with spatially-extended light source, where the resolution is normally deteriorated by the insufficient spatial coherence of the illumination. In our LIHM setup, a light-emitting diode (LED), which was a spatially-extended light source, directly illuminated the sample, and the in-line hologram were recorded by a CMOS imaging sensor located behind the sample. In our holographic reconstruction process, the in-line hologram was first deconvoled with a properly resized image of the LED illumination area, and then back-propagated with scalar diffraction formula to reconstruct the sample image. We studied the hologram forming process and showed that the additional deconvolution process besides normal scalar diffraction reconstruction in LIHM can effectively enhance the imaging resolution. The resolution enhancements capability was calibrated by numerical simulations and imaging experiments with the U.S. air force target as the sample. We also used our LIHM to image the wing of a green lacewing to further demonstrate the capability of our methods for practical imaging applications. Our methods provide a way for LIHM to achieve satisfactory resolution with less stringent requirement for spatial coherence of the source and could reduce the cost for compact imaging system.

Lensless in-line holographic microscope with Talbot grating illumination.

We have developed a wide field-of-view lensless in-line holographic microscope (LIHM) capable of acquiring microscopic images with a compact design. In our imaging system, a Ronchi grating was illuminated by a collimated laser beam to generate a Talbot self-imaging grating illumination on the sample, and the in-line holograms were recorded by a CMOS imaging sensor behind the sample. An iterative reconstruction algorithm was then applied to reconstruct the sample image while eliminating the twin-image background that appears in traditional in-line holography. In the algorithm, the dark areas of the illumination grating were used as a known constraint to define the sample support that led to convergence of the iteration. The whole-sample image can be acquired by laterally shifting the grating. We demonstrated the performance of our iteration algorithm and imaging system by successfully acquiring images of polystyrene microspheres with 5 µm diameter and the wing of a green lacewing.