Development and implementation of quadratically distorted (QD) grating and grisms system for 4D multi-colour microscopy imaging (MCMI)
The recent emergence of super-resolution microscopy imaging techniques has surpassed the diffraction limit to improve image resolution. Contrary to the breakthroughs of spatial resolution, high temporal resolution remains a challenge. This dissertation demonstrates a simple, on axis, 4D (3D + time) multi-colour microscopy imaging (MCMI) technology that delivers simultaneous 3D broadband imaging over cellular volumes, which is especially applicable to the real-time imaging of fast moving biospecimens. Quadratically distorted (QD) grating, in the form of an off axis-Fresnel zone plate, images multiple object planes simultaneously on a single image plane. A delicate mathematical model of 2D QD grating has been established and implemented in the design and optimization of QD grating. Grism, a blazed grating and prism combination, achieves chromatic control in the 4D multi-plane imaging. A pair of grisms, whose separation can be varied, provide a collimated beam with a tuneable chromatic shear from a collimated polychromatic input. The optical system based on QD grating and grisms has been simply appended to the camera port of a commercial microscope, and a few bioimaging tests have been performed, i.e. the 4D chromatically corrected imaging of fluorescence microspheres, MCF-7 and HeLa cells. Further investigation of bioimaging problems is still in progress.