Illuminating organelle dynamics through highly scattering bone: intravital wavefront optimized structured illumination imaging

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Mortensen 1706916This proposal describes a range of optical designs and engineering tasks to create an optimized microscope for use in deep tissue imaging. If successful, the proposed work will extend super-resolution structured illumination microscopy substantially and will provide a significant biological advance for tracking and monitoring engineered stem cells in deep tissues. In vivo optical imaging at depth with cellular level resolution is a major challenge to the clinical translation of biophotonic technology and the proposed resolution and depth of penetration metrics would be very beneficial to the field. The PIs support a diverse group of trainees (including high school students) and will develop two new courses on microscopy for biologists and biology for microscopists.The proposed research will benefit a range of fields including the skeletal biology, hematopoietic and neuroscience communities. To enable sub-diffraction microscopy in living bone marrow, three objectives will be pursued: 1.) A dedicated 2-photon microscope will be designed and built with a deformable mirror and digital micromirror device. 2.) The wavefront shaping 2-photon scanning microscope will be enhanced for live 2-photon structured illumination imaging resulting in 150nm lateral resolution. 3.) The instrument will be tested and validated by intravital imaging of mitochondria dynamics in Mesenchymal Stem Cells (MSCs) in mouse bone.