Detail of Figure3_701GOERGCaMP6fLckRCaMP2HeLax60x1His1uM



Project
Title
Time-lapse imaging of HeLa cells expressing Lck-RCaMP2 and OER-GCaMP6f.
Description
Time-lapse imaging of HeLa cells expressing Lck-RCaMP2 and OER-GCaMP6f.
Release, Updated
2022-11-23,
2023-02-16
Errata in Project
2023/02/16 The expression "msec" changed to "millisecond"
License
CC BY
Kind
Image data based on NA
File Formats
.stk
Data size
250.6 MB

Organism
Homo sapiens ( NCBI:txid9606 )
Strain(s)
-
Cell Line
HeLa cell ( CLO_0003684 )
Protein names
RCaMP2, GCaMP6f

Datatype
NA
Molecular Function (MF)
Biological Process (BP)
calcium ion signaling ( GO:0019722 )
Cellular Component (CC)
plasma membrane ( GO:0005886 )
Biological Imaging Method
time lapse microscopy ( Fbbi:00000249 )
X scale
0.433 micrometer/pixel
Y scale
0.433 micrometer/pixel
Z scale
-
T scale
100 milliseconds of time interval

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

Summary of Methods
See details in Bannai H et. al. J Vis Exp. 2019 Mar 22;(145).
Related paper(s)

Hiroko Bannai, Matsumi Hirose, Fumihiro Niwa, Katsuhiko Mikoshiba (2019) Dissection of Local Ca2+ Signals in Cultured Cells by Membrane-targeted Ca2+ Indicators., Journal of visualized experiments : JoVE, Number 145

Published in 2019 Mar 22 (Electronic publication in March 22, 2019, midnight )

(Abstract) Calcium ion (Ca(2+)) is a universal intracellular messenger molecule that drives multiple signaling pathways, leading to diverse biological outputs. The coordination of two Ca(2+) signal sources-"Ca(2+) influx" from outside the cell and "Ca(2+) release" from the intracellular Ca2+ store endoplasmic reticulum (ER)-is considered to underlie the diverse spatio-temporal patterns of Ca(2+) signals that cause multiple biological functions in cells. The purpose of this protocol is to describe a new Ca(2+) imaging method that enables monitoring of the very moment of "Ca(2+) influx" and "Ca(2+) release". OER-GCaMP6f is a genetically encoded Ca(2+) indicator (GECI) comprising GCaMP6f, which is targeted to the ER outer membrane. OER-GCaMP6f can monitor Ca(2+) release at a higher temporal resolution than conventional GCaMP6f. Combined with plasma membrane-targeted GECIs, the spatio-temporal Ca(2+) signal pattern can be described at a subcellular resolution. The subcellular-targeted Ca(2+) indicators described here are, in principle, available for all cell types, even for the in vivo imaging of Caenorhabditis elegans neurons. In this protocol, we introduce Ca(2+) imaging in cells from cell lines, neurons, and glial cells in dissociated primary cultures, and describe the preparation of frozen stock of rat cortical neurons.
(MeSH Terms)

Contact
Hiroko Bannai , RIKEN , RIKEN Center for Brain Science , Laboratory for Developmental Neurobiology
Contributors

OMERO Dataset
OMERO Project
Source