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Imaging Mitochondrial Calcium Fluxes with Fluorescent Probes and Single- or Two-Photon Confocal Microscopy

Davidson, SM; Duchen, MR; (2018) Imaging Mitochondrial Calcium Fluxes with Fluorescent Probes and Single- or Two-Photon Confocal Microscopy. Methods in Molecular Biology , 1782 pp. 171-186. 10.1007/978-1-4939-7831-1_10. Green open access

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Abstract

The concentration of calcium ions in the mitochondria has been shown to affect its function, modulating respiratory activity at low levels and causing lethal damage at high concentrations. The rhodamine series of dyes can be used to measure mitochondrial calcium concentration, but the reliability of measurements depends upon correct partitioning of the dye within to the mitochondria. Methods are described to aid verification and quantification of the mitochondrial calcium concentration using single- or two-photon confocal microscopy. The method of linear unmixing to separate fluorescent signals based on either differing excitation or emission spectra is outlined and for the purposes of illustration is applied to the separation of rhod-2 signals originating from the dye within the mitochondria and nucleoli.

Type: Article
Title: Imaging Mitochondrial Calcium Fluxes with Fluorescent Probes and Single- or Two-Photon Confocal Microscopy
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1007/978-1-4939-7831-1_10
Publisher version: http://dx.doi.org/10.1007/978-1-4939-7831-1_10
Language: English
Additional information: This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions.
Keywords: Calcium, Confocal microscopy, Mitochondria, Multiphoton microscopy, Rhod-2
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Cell and Developmental Biology
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Pop Health Sciences > Institute of Cardiovascular Science
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Pop Health Sciences > Institute of Cardiovascular Science > Pre-clinical and Fundamental Science
URI: https://discovery.ucl.ac.uk/id/eprint/10052217
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