Laser-based wide-field coherent imaging methods suffer from low image contrast due to the speckle noise as well as poor lateral resolution using collimated illumination. To improve the image contrast and spatial resolution for label-free cell imaging applications, we propose a new dynamic speckle illumination scheme using perfect optical vortex (POV) beams that can provide finer speckles with more uniform distributions. The low spatial coherence from the POV speckle field has significantly improved the signal-to-noise ratio (SNR) and the image contrast, thus contributing to a high spatial resolution that matches the diffraction limit in our cell imaging experiments. Importantly, the depth-resolved imaging capability has been realized which has allowed us to visualize fine subcellular structures at different focal planes.
We have recently demonstrated a high throughput three-dimensional (3D) image flow cytometry method, in which a machine-learning algorithm is used to retrieve the 3D refractive index maps of cells from one angle-multiplexing interferogram. Using this system, we have imaged flowing red blood cells and NIH/3T3 cells with a throughput of more than < 10,000 volumes/second. To further demonstrate its potential on cell phenotyping for clinical testing, we plan to apply this platform to image large populations of various cell types and extracting their morphological and biophysical parameters.
The development of high throughput three-dimensional (3D) microscopic imaging technique is important for studying cell physiology and early-stage disease diagnoses. Here we propose and demonstrate a digital micromirror device (DMD) based angle-multiplexed high-speed optical diffraction tomography (ODT) technique. Using this ODT technique, we have achieved 3D imaging of cells at over 600 tomogram/second speed, which is 10-100 times faster than current ODTbased 3D cell imaging techniques. We envision that this high-speed ODT system will enable many cutting-edge biomedical applications, such as capturing millisecond scale cell dynamics in 3D space and high throughput 3D imaging of large cell populations.
We propose and demonstrate a high sensitivity common-path quantitative phase microscopy (QPM) technique that can be used to detect nanoscale dynamics with millisecond temporal resolution. Our system is based on a transmission-mode diffraction phase microscope that is implemented with a high electron well-depth camera to reduce the phase noise. Our current system can achieve ~0.1 mrad temporal phase sensitivity, which is one order of magnitude better over most current QPM systems. Our system can be potentially used for observing morphological changes of cells and probing subnanometer membrane dynamics with millisecond temporal resolution.
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