Timelapse images and BDML file for quantitative information about the extracellular signal-regulated kinase (ERK) biosensor activity in HeLa cells, or 4T1 cancer cells or the transgenic mice
Description
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Relase date
2021-09-30
Updated date
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License
CC BY
Kind
Image data
based on Experiment
Number of Datasets
13
( Image datasets: 13,
Quantitative data datasets: 0 )
0.867 micrometer/pixel, 0.662 micrometer/pixel, Arbitrary unit by manual variable magnification.
Y scale
0.867 micrometer/pixel, 0.662 micrometer/pixel, Arbitrary unit by manual variable magnification.
Z scale
1 micrometer/slice, NA
T scale
4 minites for each time interval, 2 minites for each time interval, 121.91 second for each time interval, 1 minite for each time interval
Image Acquisition
Experiment type
-
Microscope type
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Acquisition mode
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Contrast method
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Microscope model
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Detector model
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Objective model
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Filter set
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Related paper(s)
Naoki Komatsu, Kenta Terai, Ayako Imanishi, Yuji Kamioka, Kenta Sumiyama, Takashi Jin, Yasushi Okada, Takeharu Nagai, Michiyuki Matsuda (2018) A platform of BRET-FRET hybrid biosensors for optogenetics, chemical screening, and in vivo imaging., Scientific reports, Volume 8, Number 1, pp. 8984
Published in 2018 Jun 12
(Electronic publication in June 12, 2018, midnight )
(Abstract) Genetically encoded biosensors based on the principle of Forster resonance energy transfer comprise two major classes: biosensors based on fluorescence resonance energy transfer (FRET) and those based on bioluminescence energy transfer (BRET). The FRET biosensors visualize signaling-molecule activity in cells or tissues with high resolution. Meanwhile, due to the low background signal, the BRET biosensors are primarily used in drug screening. Here, we report a protocol to transform intramolecular FRET biosensors to BRET-FRET hybrid biosensors called hyBRET biosensors. The hyBRET biosensors retain all properties of the prototype FRET biosensors and also work as BRET biosensors with dynamic ranges comparable to the prototype FRET biosensors. The hyBRET biosensors are compatible with optogenetics, luminescence microplate reader assays, and non-invasive whole-body imaging of xenograft and transgenic mice. This simple protocol will expand the use of FRET biosensors and enable visualization of the multiscale dynamics of cell signaling in live animals.
Contact
Michiyuki Matsuda
, Kyoto University
, Graduate School of Biostudies
, Laboratory of Bioimaging and Cell Signaling