Calcium measurement methods

Abstract

Calcium handling by mitochondria is important both because mitochondria can shape the cytosolic Ca2+ signals and because changes in mitochondrial Ca2+ concentration (Ca2+ physiological functions such as respiration or programmed cell death. Accurate measurements of Ca2+ require selective location of the Ca2+ M protein probes to the mitochondrial matrix. Aequorins are very adequate as Ca2+ allow molecular engineering for targeting or for changing the Ca2+ for measurements; (3) Ca2+ buffering is small; (4) have a very steep Ca2+ dynamic range, which makes them ideal for detecting and quantifying Ca2+ probe inside mitochondria and this is best achieved by targeting probes because: (1) they are important for controlling M affinity; (2) do not require irradiation -dependence and a very wide 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.