We made fluorescence multi-scale imager including 1P, 2P, light-sheet, microscope, and on-chip sensors for visualization of XYZT single cell dynamics in vivo. We integrated five microscope for systematic evaluation of living animals.
First visualization system is super-resolution imaging based on non-linear optics, with X- (resonance), Y-(galvano), and Z-(piezo) axis scanning. Real-time, multi-color XYZT multi-photon imaging enabled us to visualize single blood cell behavior in vessels and stroma. Spatial and time resolution was improved by pattern illumination. We analyzed thrombus formation, and inflammatory responses under microscope. We developed thrombus formation animal models, and elucidated cellular mechanisms of cardiovascular diseases. We directly manipulated cells by photo-chemical reactions, and two photon lasers to induce and observe thrombotic reactions. Using this system, we elucidated contribution of endothelial injuries to thrombus formation.
Second, high-resolutions and broad-imaging field was simultaneously enabled using 8K CMOS technologies, and 1P spinning disk confocal. We visualized whole organs and single cell in one image, and revealed complicated cell-cell interactions networks in single view. 8K, 60fps, and multi-color imaging visualized single platelet dynamics and tissue structural changes in single image.
Third, we performed light-sheet imaging for living mice, and enabled high-speed observation of living animals.
Fourth, macro imaging system for awake mice, pigs and humans was also developed, and free behavior monitoring revealed the tight association between metabolism and vascular reactions during daily life. Bioluminescent and fluorescent imaging from body surface using CMOS camera, image intensifier, and macro-lens enabled us to visualize cellular dynamics without anesthesia.
Last, wearable, implantable, and minimized devices for non-invasive recording were also developed using lens-less and on-chip technologies. We utilized SCMOS, micro-lens array, and LED illumination technologies.
In sum, we developed multi-scale imaging system which can evaluate cellular networks dysregulations in diseased conditions.
To cite this abstract:Satoshi Nishimura; Multi-modal in vivo visualization of single cell dynamics by 1P, 2P, light-sheet, and on-chip technologies.. The 16th European Microscopy Congress, Lyon, France. https://emc-proceedings.com/abstract/multi-modal-in-vivo-visualization-of-single-cell-dynamics-by-1p-2p-light-sheet-and-on-chip-technologies/. Accessed: May 26, 2020
EMC Abstracts - https://emc-proceedings.com/abstract/multi-modal-in-vivo-visualization-of-single-cell-dynamics-by-1p-2p-light-sheet-and-on-chip-technologies/