eBook: Advanced Tools Transforming Neurology Research - 19

Fluorescence-based Calcium
Mobilization Assay with the
CELENA® X High Content
Imaging System
An optimized analysis pipeline to evaluate ATP-induced
intracellular calcium flux
Key features
* High throughput screening of membrane
potentials with the CELENA® X automated
microscope system.
* ATP inducible Fluo-8® AM esters into live cells
for Kinetic live cell imaging.
* The optimized analysis pipeline to evaluate
ATP-induced intracellular calcium flux.
Introduction
Calcium is an essential element found in all living
organisms and is necessary for many functions
at the cellular level. Calcium migrates across cell
membranes and permits the activation and inhibition
of a variety of enzymes through embedded
calcium ion channels.1-3
These calcium ions are
responsible for facilitating muscle relaxation and
contraction, as well as, vascular contraction and
vasodilation, neuronal signal transmission, and
hormone secretion.4-6
This assessment evaluates
the changes of intracellular calcium flux conditions
on bovine aortic endothelial cells (BAEC) by adding
ATP agonist.7
The Fluo-8® AM, a acetoxymethyl ester
of fluorescent indicator dye, is hydrolyzed intracellularly
in the presence of calcium ions. Thus, the
combinations of Fluo-8® AM and ATP agonist were
used to detect changes in intracellular calcium efflux
levels in the live cells. A 3-minute time-lapse
was set for monitoring the BAEC cellular calcium
efflux activated by the ATP stimulant forced the
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