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Isotonic and minimally invasive optical clearing media for live cell imaging ex vivo and in vivo

Inagaki, S.; Nakagawa-Tamagawa, N.; Huynh, N.; Kambe, Y.; Yagasaki, R.; Manita, S.; Fujimoto, S.; Noda, T.; Mori, M.; Teranishi, A.; Takeshima, H.; Naito, Y.; Yokoyama, T.; Sakamoto, M.; Hayashi, K.; Kitamura, K.; Tagawa, Y.; Okuda, S.; Sato, T.; Imai, T.

2024-09-15 neuroscience
10.1101/2024.09.13.612584 bioRxiv
Show abstract

Tissue clearing has been widely used for fluorescence imaging of fixed tissues, but not for live tissues due to its toxicity. Here we develop minimally invasive optical clearing media for fluorescence imaging of live mammalian tissues. Light scattering is minimized by adding spherical polymers with low osmolarity to the extracellular medium. A clearing medium containing bovine serum albumin (SeeDB-Live) is minimally invasive to live cells, allowing for structural and functional imaging of live tissues, such as spheroids, organoids, acute brain slices, and the mouse brain in vivo. SeeDB-Live minimally affects the electrophysiological properties and sensory responses of neurons. We demonstrate its utility for widefield imaging of subcellular voltage dynamics, such as backpropagating action potentials, in acute brain slices. We also utilize SeeDB-Live for widefield voltage imaging of dozens of dendrites in vivo, demonstrating population dynamics. Thus, SeeDB-Live expands the scale and modalities of fluorescence imaging of live mammalian tissues.

Published in Nature Methods (predicted rank #2) · training set

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