High-throughput 3D Image Cytometer

One of my major contributions is towards 3D image cytometry. Image cytometry provides a wealth of information about the native state of cells and tissues. The ability to perform fast 3D measurements further enhances the information throughput of image cytometers. I have worked on 3D cytometers based on structured light illumination, and multiphoton temporal-focusing wide-field imaging. First, we built a high-throughput image cytometer based on structured light illumination and remote focusing [a]. The instrument was capable of imaging ~ 800 cells per second in 3D, which is an order of magnitude faster than the current state of the art. Moreover, we developed an arsenal of custom image analysis algorithms for large data sets generated by this platform. Together, with the instrument and algorithms, we demonstrated rare cellular event detection (up to 1 in 100,000) in large cell populations. Second, we designed a novel deep tissue 3D imaging microscope based on wide-field temporal-focusing imaging. Temporal focusing is an ideal technique for depth-resolved wide-field excitation deep inside living tissue. However, the emission light is usually subjected to scattering, degrading the image quality. The new architecture contained a digital mirror device (DMD) that is imaged onto the temporal focusing plane. The DMD projected predefined structured patterns that encoded spatial features in the focal plane. The images were then recorded on a CMOS camera and decoded using compressive sensing reconstruction algorithms.

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