Spectral Domain Phase Microscopy: A novel investigative tool for cellular dynamics

Optical coherence tomography (OCT) is an optical interferometry technique that produces high-resolution images of tissue microstructure. The emergence of a new generation of super-broadband sources has facilitated construction of OCT systems with micron-scale resolution. Noninvasive and easily adaptable, OCT is an ideal imaging modality to complement the variety of tools already available to geneticists and cell biologists. Arguably, micron-resolution is sufficient for viewing large-scale cellular motions, however future advances in imaging subcellular functions will demand resolution at the sub-micron resolution scale. Currently, OCT is incapable of providing the resolution performance required to resolve sub-micron dynamic events.;Phase-sensitive detection has a long history as means for generating contrast in transparent samples. This dissertation describes the development and deployment of Spectral Domain Phase Microscopy (SDPM) for biological imaging. As a phase-sensitive functional derivative of OCT, the axial resolution of SDPM remains linked to that of contemporary OCT systems, but the technique enables sub-micron resolution of dynamic cellular phenomena. A microscope-adapted SDPM system for three-dimensional imaging was constructed to enable simultaneous SDPM and bright field imaging. Biological investigations of single cells examined such dynamic phenomena as locomotion, cardiac contraction and viscoelastic properties of the cytoskeleton. Mathematical expressions are derived to describe the underlying theory for phase washout and phase corruption, two sources of phase error that were encountered in the course of this work.