| This dissertation describes the development and application of multimodal optical imaging methods for providing enhanced visualization of living skin. In vivo imaging of skin is of great interest for research applications to study fundamental aspects of skin biology as well as for clinical applications to potentially enable non-invasive diagnosis of skin diseases. The methods presented are based on a multimodal microscope that integrates optical coherence and multiphoton microscopy. The development of the integrated microscope was focused on enabling sufficient performance of both modalities for in vivo imaging applications. This included the design of a dual-spectrum laser source that meets the requirements needed by the different imaging modalities as well an image reconstruction algorithm for optical coherence microscopy that corrects curvature aberrations and significantly reduces computational complexity. The multimodal contrast is capable of providing a view of many different components of living skin, including macroscopic and microscopic structure, microvasculature, collagen networks, single cells and single cell nuclei. In addition to multimodal imaging, technical challenges associated with long-term tracking of the same skin site in vivo are addressed through the development of a non-rigid registration algorithm. For this dissertation, these imaging methods were applied in several studies that were conducted in both human and mouse skin. Emphasis was placed on the application of these imaging methods for studying fundamental biology in mouse skin, including time-lapse imaging of wound healing, skin grafting and single-cell dynamics. In addition the imaging methods were applied to studying the potential toxicity of nanoparticles following skin exposure. The imaging methods presented in this dissertation have great potential for facilitating a deeper understanding of fundamental skin biology and for enabling non-invasive assessment of diseases. |
