Optical nanoscopy has changed the “seeing is believing” paradigm. This was achieved within a limited field of view (FOV~100µm2) and required fluorescent markers. Large-FOV high-throughput live unimpaired cell imaging is crucial fo...
Optical nanoscopy has changed the “seeing is believing” paradigm. This was achieved within a limited field of view (FOV~100µm2) and required fluorescent markers. Large-FOV high-throughput live unimpaired cell imaging is crucial for biology and biomedicine. Hence, improving the space bandwidth product (SBP) using time-consuming scanning stitching is not a good solution. Lensless holographic microscopy (LHM) inherently bypasses FOV limitations by using full sensor-size hologram reconstruction for label-free object information retrieval. Its major limitation, not yet addressed, is its low lateral (~1µm) and axial (~3µm) resolution. I will overcome this fundamental problem by pioneering deep UV (DUV) lensless holotomographic nanoscopy (LHN) as a simple and compact device easily operated inside the cell chamber or outside the laboratory (in contrast to lens-based systems). Owing to DNA damage, DUV is used to sterilize and never image bio-samples. This paradigm will be shifted to provide a breakthrough 10 giga pixel SBP via a low-dose DUV optical-elements-free (no cost, no radiation loss) lensless setup with a worlds-first full-angle tomographic scenario, numerical aperture > 1, and a new class of reconstruction algorithms to decrease the effective pixel (to 100 nm) and remove background noise. I will use LHN to enable the discovery of a new mechanistic understanding of extracellular vesicles expression and intake within large live cell cultures with single-vesicle resolution. EVs, nanosized lipid spheres released by virtually every cell type, are currently emerging as novel disease biomarkers and drug nanovehicles. LHN is a new research field that inherently makes this a high-risk project, but the potential gains are also high as a new era of simple ultrahigh SBP nanoimaging might be opened. The proposed multidisciplinary project calls for near-unique expertise in computational microscopy and digital holography, which I acquired in cooperation with international leaders.ver más
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