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During the action potential of excitable cells, intracellular calcium levels transiently increase due to release of calcium ions from the extracellular environment and sarcoplasmic reticulum into the cytoplasm. Ncardia can monitor this fluctuation in intracellular calcium levels using two methods:
Calcium-sensitive dyes that exhibit changes in fluorescence upon binding to calcium ions are accurate and rapid indicators of calcium changes in the intracellular environment. When added to cell culture plates, they are absorbed into the cytoplasm, allowing real-time monitoring of changes in intracellular calcium concentrations. Ncardia's calcium signaling service allows simultaneous dispensing of test compounds and real-time measurement of intracellular calcium transients.
Genetically encoded calcium indicators, such as GCaMP, are also powerful tools for measuring calcium signals. When introduced into excitable cells, genetically encoded calcium indicators enable monitoring of changes in intracellular calcium concentrations in a non-invasive long-term way, making them highly applicable for studying chronic cytotoxic effects of your compounds.
In this case study, we assessed the effects of a set of known cardioactive compounds on Pluricyte Cardiomyocytes using the FLIPR Tetra High-Throughput Cellular Screening System (Molecular Devices). Calcium transients were recorded and used to analyze specific cardioactive compound response parameters, including peak frequency, peak amplitudes, and average peak width. Below an example is shown of the effect of different reference compounds.
Click here to read the full case study: Compound effects on calcium transients in Pluricyte Cardiomyocytes
Effects of Reference Compounds on Calcium Transients in Pluricyte Cardiomyocytes
The negative control, 0.25% DMSO, shows a representative calcium transient pattern of untreated Pluricyte Cardiomyocytes. Treatment with cardioactive compounds, known to directly or indirectly affect calcium transients, demonstrated compound-specific effects:
The calcium channel agonist, Bay K8644, at a concentration of 1.26 µM caused increased peak amplitudes, increased peak widths, and reduced peak frequency. By blocking L-type calcium channels with 19 nM nifedipine, the cells show reduced calcium transient peak amplitudes. Treatment with the β- adrenergic receptor agonist, isoprenaline (500 nM), induced increased peak frequency, and decreased peak width.
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