Imaging intraorganellar Ca2+ at subcellular resolution using CEPIA
Citations Over TimeTop 1% of 2014 papers
Abstract
The endoplasmic reticulum (ER) and mitochondria accumulate Ca(2+) within their lumens to regulate numerous cell functions. However, determining the dynamics of intraorganellar Ca(2+) has proven to be difficult. Here we describe a family of genetically encoded Ca(2+) indicators, named calcium-measuring organelle-entrapped protein indicators (CEPIA), which can be utilized for intraorganellar Ca(2+) imaging. CEPIA, which emit green, red or blue/green fluorescence, are engineered to bind Ca(2+) at intraorganellar Ca(2+) concentrations. They can be targeted to different organelles and may be used alongside other fluorescent molecular markers, expanding the range of cell functions that can be simultaneously analysed. The spatiotemporal resolution of CEPIA makes it possible to resolve Ca(2+) import into individual mitochondria while simultaneously measuring ER and cytosolic Ca(2+). We have used these imaging capabilities to reveal differential Ca(2+) handling in individual mitochondria. CEPIA imaging is a useful new tool to further the understanding of organellar functions.
Related Papers
- → Calcium signaling and apoptosis(2003)468 cited
- → High speed optically sectioned fluorescence lifetime imaging permits study of live cell signaling events(2007)84 cited
- → Imaging fluorescence lifetime heterogeneity applied to GFP‐tagged MHC protein at an immunological synapse*(2005)68 cited
- → Changes to lipid droplet configuration in mCMV-infected fibroblasts: live cell imaging with simultaneous CARS and two-photon fluorescence microscopy(2011)11 cited
- → Implementation of miniaturized modular-array fluorescence microscopy for long-term live-cell imaging(2023)