A Multimodal Adaptive Optical Microscope For In Vivo Imaging from Molecules to Organisms

Fu, T.-M.; Liu, G.; Milkie, D. E.; Ruan, X.; Gorlitz, F.; Shi, Y.; Ferro, V.; Divekar, N. S.; Wang, W.; York, H. M.; Kilic, V.; Mueller, M.; Liang, Y.; Daugird, T. A.; Gacha-Garay, M. J.; Larkin, K. A.; Adikes, R. C.; Harrison, N.; Shirazinejad, C.; Williams, S.; Nourse, J. L.; Sheu, S.-H.; Gao, L.; Li, T.; Mondal, C.; Achour, K.; Hercule, W.; Stabley, D.; Emmerich, K.; Dong, P.; Drubin, D.; Liu, Z. J.; Clapham, D.; Mumm, J. S.; Koyama, M.; Killilea, A.; Bravo-Cordero, J. J.; Keene, C. D.; Luo, L.; Kirchhausen, T.; Pathak, M. M.; Arumugam, S.; Nunez, J. K.; Gao, R. G.; Matus, D. Q.; Martin, B.

doi:10.1101/2025.06.02.657494

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bioRxiv · 10.1101/2025.06.02.657494

A Multimodal Adaptive Optical Microscope For In Vivo Imaging from Molecules to Organisms

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Abstract

Understanding biological systems requires observing features and processes across vast spatial and temporal scales, spanning nanometers to centimeters and milliseconds to days, often using multiple imaging modalities within complex native microenvironments. Yet, achieving this comprehensive view is challenging because microscopes optimized for specific tasks typically lack versatility due to inherent optical and sample handling trade-offs, and frequently suffer performance degradation from sample-induced optical aberrations in multicellular contexts. Here, we present MOSAIC, a reconfigurable microscope that integrates multiple advanced imaging techniques including light-sheet, label-free, super-resolution, and multi-photon, all equipped with adaptive optics. MOSAIC enables non-invasive imaging of subcellular dynamics in both cultured cells and live multicellular organisms, nanoscale mapping of molecular architectures across millimeter-scale expanded tissues, and structural/functional neural imaging within live mice. MOSAIC facilitates correlative studies across biological scales within the same specimen, providing an integrated platform for broad biological investigation.

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Fu, T.-M., Liu, G., Milkie, D. E., Ruan, X., Gorlitz, F., Shi, Y., Ferro, V., Divekar, N. S., Wang, W., York, H. M., Kilic, V., Mueller, M., Liang, Y., Daugird, T. A., Gacha-Garay, M. J., Larkin, K. A., Adikes, R. C., Harrison, N., Shirazinejad, C., Williams, S., Nourse, J. L., Sheu, S.-H., Gao, L., Li, T., Mondal, C., Achour, K., Hercule, W., Stabley, D., Emmerich, K., Dong, P., Drubin, D., Liu, Z. J., Clapham, D., Mumm, J. S., Koyama, M., Killilea, A., Bravo-Cordero, J. J., Keene, C. D., Luo, L., Kirchhausen, T., Pathak, M. M., Arumugam, S., Nunez, J. K., Gao, R. G., Matus, D. Q., Martin, B.. 2025-06-16. A Multimodal Adaptive Optical Microscope For In Vivo Imaging from Molecules to Organisms. https://doi.org/10.1101/2025.06.02.657494

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