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- W2480087180 abstract "Calcium handling by mitochondria is important both because mitochondria can shape the cytosolic Ca2+ signals and because changes in mitochondrial Ca2+ concentration ([Ca2+]M) are important for controlling physiological functions such as respiration or programmed cell death. Accurate measurements of [Ca2+]M require selective location of the Ca2+ probe inside mitochondria and this is best achieved by targeting protein probes to the mitochondrial matrix. Aequorins are very adequate as Ca2+probes because: (1) they allow molecular engineering for targeting or for changing the Ca2+ affinity; (2) do not require irradiation for measurements; (3) Ca2+ buffering is small; (4) have a very steep Ca2+-dependence and a very wide dynamic range, which makes them ideal for detecting and quantifying Ca2+ microdomains. Consumption and low light output are some of its drawbacks that make calcium imaging a hard task. Here, we describe a procedure that overcomes these disadvantages by combining herpes simplex virus type 1(HSV-1)-based expression of targeted aequorins with photon-counting imaging. This methodology allows real-time resolution of changes of [Ca2+]M by photon counting imaging at the single-cell level. Since HSV virus is neurotrophic, the method is adequate for measuring [Ca2+]M in living neurons." @default.
- W2480087180 created "2016-08-23" @default.
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- W2480087180 date "2009-11-10" @default.
- W2480087180 modified "2023-10-18" @default.
- W2480087180 title "Ca2+ Imaging of Intracellular Organelles: Mitochondria" @default.
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- W2480087180 doi "https://doi.org/10.1007/978-1-60761-476-0_9" @default.
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