Multi-focus microscopy for aberration-corrected multi-color 3D imaging

Due to the classical conflict between spatial and temporal resolution, microscopy studies of fast events in living samples are often performed in 2D even when 3D imaging would be desirable and could provide new insights to biological function. This dissertation describes an instant 3D imaging system - a multi-focus microscope (MFM) - which provides high- resolution, aberration-corrected, multi-color fluorescence images of multiple focal planes simultaneously. Forming an instant focal series eliminates the need for multiple camera exposures and mechanical refocusing, allowing 3D imaging limited only by sample signal strength and the camera read-out rate for a single frame. A module containing the MFM optical components can easily be appended to the camera port of a commercial wide-field microscope. The excellent resolution and sensitivity of MFM is demonstrated on two different 3D biological imaging problems; neuronal imaging in the entire C.elegans embryo and mRNA imaging in cultured mammalian cells