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Imaging H2O2 microdomains in receptor tyrosine kinases signaling.
- Source :
-
Methods in enzymology [Methods Enzymol] 2013; Vol. 526, pp. 175-87. - Publication Year :
- 2013
-
Abstract
- HyPer, a ratiometric genetically encoded fluorescent sensor, is a popular tool for intracellular hydrogen peroxide detection. When expressed in cultured cells, the freely diffusing version of the sensor (HyPer-cyto) detects temporal patterns of H2O2 generation. However, rapid diffusion of the probe within the nucleocytoplasmic compartment averages the H2O2 signal even in cases of local oxidant production. Consequently, we immobilized the sensor within specific subcellular compartments allowing it to monitor local increases in H2O2. Here, we provide a protocol of ratiometric imaging and ImageJ-based quantification of H2O2 microdomains produced by cells upon physiological stimulation.<br /> (Copyright © 2013 Elsevier Inc. All rights reserved.)
- Subjects :
- Animals
Cell Line
Fluorescent Dyes metabolism
Humans
Hydrogen Peroxide metabolism
Image Processing, Computer-Assisted methods
Microscopy, Confocal methods
Microscopy, Fluorescence methods
Transfection methods
Fluorescent Dyes analysis
Hydrogen Peroxide analysis
Optical Imaging methods
Receptor Protein-Tyrosine Kinases metabolism
Signal Transduction
Subjects
Details
- Language :
- English
- ISSN :
- 1557-7988
- Volume :
- 526
- Database :
- MEDLINE
- Journal :
- Methods in enzymology
- Publication Type :
- Academic Journal
- Accession number :
- 23791101
- Full Text :
- https://doi.org/10.1016/B978-0-12-405883-5.00011-9