We are proposing to develop the polarized light field microscope for analyzing the architectural dynamics inside living cells, tissues and cell-free model systems. The instrument combines comprehensive polarization analysis and 3D imaging of microscopic specimens in a single snapshot using the new approach of light field imaging. The project includes the development of the hardware, software and algorithms required for capturing polarized light field images and their tomographic reconstruction into 3-dimensional maps of birefringence, diattenuation and polarized fluorescence. The instrument development is guided by several application projects including the remodeling of the extracellular collagen matrix by fibroblast cells, the spatial coordination in the assembly of higher order septin structures inside eukaryotic cells and the origins of chromosome malorientations and their repair in the meiotic spindle of spermatocytes. Information about the technology, including software components and instrument design that are developed under this grant, will be made available for users and developers on an established web portal called OpenPolScope.org.
The microscopy instrumentation developed under this grant will have applications in medical research and clinical settings. Furthermore, the cell biological application projects that will be forthcoming under this grant will have broad implications for our understanding of cell division and cellular remodeling of tissue, which are fundamental processes in health and disease.
|Mehta, Shalin B; McQuilken, Molly; La Riviere, Patrick J et al. (2016) Dissection of molecular assembly dynamics by tracking orientation and position of single molecules in live cells. Proc Natl Acad Sci U S A 113:E6352-E6361|