Detail of ExtFig9def_InVivoVoltage_anesth_ctrl


Project
SSBD:Repository
Title
Time-lapse images of Voltron2-ST labelled L2/3 neurons in S1 of anesthetized mice before clearing with SeeDB-Live
Description
Time-lapse images of Voltron2-ST labelled neurons in layer 2/3 of somatosensory cortex of anesthetized mouse before clearing with SeeDB-Live. Voltron2-ST, a fast and sensitive chemigenetic voltage indicator was introduced to layer 2/3 neurons by in utero electroporation. Cell body of the neurons at 120-150 micrometer depth of the brain was imaged by epifluorescence microscopy. The brain surface was perfused with ACSF-HEPES during imaging.
Release, Updated
2026-08-19
License
CC BY 4.0
Kind
Image data
File Formats
.tif
Data size
1.2 GB

Organism
Mus musculus ( NCBI:txid10090 )
Strain(s)
-
Cell Line
-

Datatype
-
Molecular Function (MF)
Biological Process (BP)
neuronal action potential propagation
Cellular Component (CC)
cell body dendrite
Biological Imaging Method
fluorescence microscopy ( Fbbi:00000246 )
X scale
2.89 micrometer
Y scale
2.89 micrometer
Z scale
-
T scale
1 milliseconds

Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
-
Microscope model
-
Detector model
-
Objective model
-
Filter set
-

Summary of Methods
Inagaki S, Nakagawa-Tamagawa N, Huynh NZ, 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, Sato TK, Imai T. Isotonic and minimally invasive optical clearing media for live cell imaging ex vivo and in vivo. Nat Methods. 2026 Apr;23(4):839-853.
Related paper(s)

Shigenori Inagaki, Nao Nakagawa-Tamagawa, Nathan Zechen Huynh, Yuki Kambe, Rei Yagasaki, Satoshi Manita, Satoshi Fujimoto, Takahiro Noda, Misato Mori, Aki Teranishi, Hikari Takeshima, Koki Ishikawa, Yuki Naitou, Tatsushi Yokoyama, Masayuki Sakamoto, Katsuhiko Hayashi, Kazuo Kitamura, Yoshiaki Tagawa, Satoru Okuda, Tatsuo K Sato, Takeshi Imai (2026) Isotonic and minimally invasive optical clearing media for live cell imaging ex vivo and in vivo., Nature methods

Published in March 12, 2026 (Electronic publication in March 12, 2026, midnight )

(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.

Contact
Shigenori Inagaki, Takeshi Imai , Kyushu University, Kyushu University , Department of Developmental Neurophysiology, Department of Developmental Neurophysiology , Department of Developmental Neurophysiology, Department of Developmental Neurophysiology
Contributors

OMERO Dataset
OMERO Project
Source