OSCR

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

Overview

Authors: Shigenori Inagaki1,2, Nao Nakagawa-Tamagawa3, Nathan Zechen Huynh4, Yuki Kambe4, Rei Yagasaki5, Satoshi Manita6, Satoshi Fujimoto1, Takahiro Noda1, Misato Mori5, Aki Teranishi5, Hikari Takeshima1, Koki Ishikawa1, Yuki Naitou7, Tatsushi Yokoyama8,9, Masayuki Sakamoto8,10, Katsuhiko Hayashi7,11,12, Kazuo Kitamura6, Yoshiaki Tagawa3, Satoru Okuda5,13, Tatsuo K Sato2,4, Takeshi Imai1,13
ORCID iDs: Shigenori Inagaki, Nao Nakagawa-Tamagawa, Nathan Zechen Huynh, Yuki Kambe, Rei Yagasaki, Satoshi Manita, Satoshi Fujimoto, Aki Teranishi, Tatsushi Yokoyama, Masayuki Sakamoto, Katsuhiko Hayashi, Kazuo Kitamura, Satoru Okuda, Takeshi Imai
13 affiliations
  1. Department of Developmental Neurophysiology, Graduate School of Medical Sciences, Kyushu University, Higashi-ku, Fukuoka, Japan
  2. FOREST, Japan Science and Technology Agency, Saitama, Japan
  3. Department of Physiology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan
  4. Department of Pharmacology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan
  5. Nano Life Science Institute, Kanazawa University, Kanazawa, Japan
  6. Department of Neurophysiology, Faculty of Medicine, University of Yamanashi, Chuo, Japan
  7. Department of Stem Cell Biology and Medicine, Graduate School of Medical Sciences, Kyushu University, Higashi-ku, Fukuoka, Japan
  8. Department of Optical Neural and Molecular Physiology, Graduate School of Biostudies, Kyoto University, Kyoto, Japan
  9. Laboratory of Optical Biomedical Science, Institute for Life and Medical Sciences, Kyoto University, Kyoto, Japan
  10. Center for Living Systems Information Science, Graduate School of Biostudies, Kyoto University, Kyoto, Japan
  11. Department of Genome Biology, Graduate School of Medicine, Osaka University, Suita, Japan
  12. Division of Reproductive Systems, Premium Research Institute for Human Metaverse Medicine (WPI-PRIMe), Osaka University, Suita, Japan
  13. CREST, Japan Science and Technology Agency, Saitama, Japan
Institutions: Kyushu University (Japan); Japan Science and Technology Agency (Japan); Kagoshima University (Japan); Kanazawa University (Japan); University of Yamanashi (Japan); Kyoto University (Japan); Institute for Life and Medical Sciences, Kyoto University (Japan); The University of Osaka (Japan)
Journal: Nature methods, volume 23, issue 4, pages 839-853
Dates: received 1 November 2024; accepted 3 February 2026; published online 12 March 2026; in print 2026
Type: Research article · Language: English
License: CC BY
Identifiers: DOI 10.1038/s41592-026-03023-y · PMID 41820664 · PMCID PMC13076227 · OpenAlex W7135088429
Open access: hybrid, a free copy (OpenAlex)
Status: code on request
Categories: human (organism), mouse (organism)
Methods: Spectral & time-frequency, Statistics, Machine learning, Evoked potentials, fMRI & imaging, Single-unit activity, calcium imaging, Smoothing, state filtering, decompositions, Connectivity
Keywords: Neuroscience, Biophysics, Mouse, Fluorescence imaging
MeSH: Optical Imaging*, Serum Albumin, Bovine*, Animals, Brain, Humans, Mice, Neurons (* major topic)
Topic: Advanced Fluorescence Microscopy Techniques (Biophysics, Biochemistry, Genetics and Molecular Biology), according to OpenAlex
Citations: cited by 7 papers (Europe PMC); 82 references in the paper
Research resources: RRID:SCR_013672, RRID:SCR_014215

Abstract

Tissue clearing has been widely used for fluorescence imaging of fixed tissues, but its application to live tissues has been limited by 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 compatible with live cells, enabling structural and functional imaging of live tissues, such as spheroids, organoids, acute brain slices and the mouse brains in vivo. SeeDB-Live minimally affects neuronal electrophysiological properties and sensory responses in vivo, and facilitates fluorescence imaging of deep cortical layers in live animals without detectable toxicity to neurons or behavior. We further demonstrate its utility to epifluorescence voltage imaging in acute brain slices and in vivo preparations. Thus, SeeDB-Live expands both the depth and modality range of fluorescence imaging in live mammalian tissues.

Reproduced under the paper's license (CC BY), from the paper cited above.

Code

The paper says that its authors' code is available on request: it was not published with the paper, so there is nothing to verify.

The paper's code and data availability statement is in the Data section.

Tracing map

A tracing map links a paper to the code its authors published: this paper has none (its code is available on request), so it has no map.

Data

No dataset and no data link were found in the paper.

Data availability

Raw image data used in this study has been deposited to the SSBD:repository (10.24631/ssbd.repos.2025.11.484)76. Detailed protocols and technical tips are described in SeeDB Resources (https://sites.google.com/site/seedbresources/). Requests for additional program codes and data generated and/or analyzed during the current study should be directed to and will be fulfilled upon reasonable request by the corresponding author.

Reproduced under the paper's license (CC BY), from the paper cited above.

Versions

The history of this record: each version stored by the harvester or made by a correction of its authors or of the maintainers of its code, and what changed in its facts. The texts of the paper (its abstract, its availability statements) are not part of it; versions that changed only those are not listed.

Version 1, 30 September 2026: the first record

Recorded: type, language, journal, volume, issue, pages, dates, 21 authors, 4 keywords, 7 MeSH terms, 79 references, 2 RRIDs.

Cite

This paper

Inagaki, S., Nakagawa-Tamagawa, N., Huynh, N. Z., Kambe, Y., Yagasaki, R., Manita, S., Fujimoto, S., Noda, T., Mori, M., Teranishi, A., Takeshima, H., Ishikawa, K., Naitou, Y., Yokoyama, T., Sakamoto, M., Hayashi, K., Kitamura, K., Tagawa, Y., Okuda, S., . . . Imai, T. (2026). Isotonic and minimally invasive optical clearing media for live cell imaging ex vivo and in vivo. Nature methods, 23(4), 839-853. https://doi.org/10.1038/s41592-026-03023-y

BibTeX

@article{inagaki2026isotonic,
author = {Inagaki, Shigenori and Nakagawa-Tamagawa, Nao and Huynh, Nathan Zechen and Kambe, Yuki and Yagasaki, Rei and Manita, Satoshi and Fujimoto, Satoshi and Noda, Takahiro and Mori, Misato and Teranishi, Aki and Takeshima, Hikari and Ishikawa, Koki and Naitou, Yuki and Yokoyama, Tatsushi and Sakamoto, Masayuki and Hayashi, Katsuhiko and Kitamura, Kazuo and Tagawa, Yoshiaki and Okuda, Satoru and Sato, Tatsuo K and Imai, Takeshi},
title = {{Isotonic and minimally invasive optical clearing media for live cell imaging ex vivo and in vivo}},
journal = {Nature methods},
year = {2026},
month = mar,
volume = {23},
number = {4},
pages = {839--853},
publisher = {Nature Portfolio},
issn = {1548-7091},
doi = {10.1038/s41592-026-03023-y},
url = {https://doi.org/10.1038/s41592-026-03023-y},
pmid = {41820664},
pmcid = {PMC13076227}
}

RIS

TY - JOUR
AU - Inagaki, Shigenori
AU - Nakagawa-Tamagawa, Nao
AU - Huynh, Nathan Zechen
AU - Kambe, Yuki
AU - Yagasaki, Rei
AU - Manita, Satoshi
AU - Fujimoto, Satoshi
AU - Noda, Takahiro
AU - Mori, Misato
AU - Teranishi, Aki
AU - Takeshima, Hikari
AU - Ishikawa, Koki
AU - Naitou, Yuki
AU - Yokoyama, Tatsushi
AU - Sakamoto, Masayuki
AU - Hayashi, Katsuhiko
AU - Kitamura, Kazuo
AU - Tagawa, Yoshiaki
AU - Okuda, Satoru
AU - Sato, Tatsuo K
AU - Imai, Takeshi
TI - Isotonic and minimally invasive optical clearing media for live cell imaging ex vivo and in vivo
T2 - Nature methods
J2 - Nat Methods
PY - 2026
DA - 2026/03/12
VL - 23
IS - 4
SP - 839
EP - 853
SN - 1548-7091
PB - Nature Portfolio
DO - 10.1038/s41592-026-03023-y
UR - https://doi.org/10.1038/s41592-026-03023-y
LA - en
ER -

CSL-JSON

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