Isotonic and minimally invasive optical clearing media for live cell imagingex vivoandin vivo

2024 · doi:10.1101/2024.09.13.612584
preprint OA: closed CC-BY-NC-4.0
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The paper develops minimally invasive optical clearing media that reduce light scattering while maintaining low osmolarity, enabling fluorescence imaging of live mammalian tissues instead of only fixed tissue. Using a bovine serum albumin–based formulation (SeeDB-Live), the authors show it supports structural and functional imaging across multiple ex vivo preparations (e.g., spheroids, organoids, acute brain slices) and in vivo mouse brain imaging, including widefield subcellular voltage dynamics such as backpropagating action potentials and population-level dendritic activity. They report that SeeDB-Live minimally affects neuronal electrophysiological properties and sensory responses, with an explicit limitation that live-tissue compatibility was assessed under their experimental conditions rather than universally across all tissue types and functions. This paper does not explicitly discuss endometriosis or adenomyosis; it was included in the corpus via a keyword match in the upstream search index.

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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.
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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. Competing Interest Statement S.I. and T.I. have filed a patent application related to SeeDB-Live. The other authors declare that they have no competing interests. Footnotes ↵12 Lead Contact.

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License: CC-BY-NC-4.0