Detail of Fig5k_awakeL5_SDL_day1


Project
SSBD:Repository
Title
Time-lapse calcium imaging of L5ET neurons in primary somatosensory cortex using SeeDB-Live
Description
Time-lape images of awake mouse cerebral cortex using SeeDB-Live. The neurons at a depth of 615 micrometer from the surface of the brain were imaged by two-photon confocal microscopy. jGCaMP8m expressed in the layer 5 extratelencephalic-projecting (L5ET) neurons were observed in the mouse that was injected AAV-jGCaMP8m-P2A-CyRFP1. The brain surface was perfused with SeeDB-Live. The mouse was imaged two times at conscutive days. This is the imaging data at the first day.
Release, Updated
2026-08-19
License
CC BY 4.0
Kind
Image data
File Formats
.tif
Data size
502.2 MB

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

Datatype
-
Molecular Function (MF)
Biological Process (BP)
calcium ion transport
Cellular Component (CC)
cell body
Biological Imaging Method
two-photon laser scanning microscopy ( Fbbi:00000254 )
X scale
1.043 micrometer
Y scale
1.043 micrometer
Z scale
-
T scale
0.169 seconds

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