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Calcium-flux assay on the FDSS/µCELL System

Calcium ions play a critical role in the electrophysiology of the heart and regulate the activities of motor proteins necessary for cardiac contraction. Direct acute drug effects on both electrophysiology and contractility can be detected by monitoring Ca2+ transients, making it a valuable tool in drug discovery and safety pharmacology assessment. The FDSS/µCell instrument (Hamamatsu) is a kinetic plate reader with an integrated dispensing head and imaging-based detector. Upon treatment with a calcium fluorescent dye or Axiogenesis’ Xpress.4U/GCAMP6f transfection reagent, real-time changes in fluorescent calcium transients can be measured in Cor.4U® or Pluricyte® Cardiomyocytes at a high-throughput (96- and 384-well plate formats).

User guide

Cor.4U® and Pluricyte® Cardiomyocytes are optimized for use on the FDSS/µCELL System

The use of Cor.4U® and Pluricyte® Cardiomyocytes in combination with the FDSS/µCell instrument has been optimized in house. Combining these technologies enables the evaluation of potential cardiotoxic effects of compounds at high-throughput (up to 384 wells), thereby supporting your decision making in the safety of drug candidates at an early stage of drug development.

You can find a step-by-step description of the analysis of fluorescent dye-based calcium transients of our hiPSC-derived cardiomyocytes using the FDSS/µCell System in our User Guides:

Application

Drug safety assessment using hiPSC-derived cardiomyocytes in combination with the FDSS/mCELL System

Case studies were performed to assess the acute effects of well-known cardioactive compounds in Cor.4U® and Pluricyte® Cardiomyocytes calcium transients using the FDSS/mCELL kinetic plate reader. Acute effects of cardioactive compounds were determined to analyze various parameters, including peak frequency, peak amplitudes, peak slopes, and average peak width duration. Below an example is shown of the effect of the reference compound, astemizole, in Cor4U® cardiomyocytes.

Cor.4U® show  reduced peak frequency and arrhythmia in response to astemizole

Astemizole is a hERG channel blocker that can induce arrhythmias at higher concentrations. Treatment of Cor.4U® cardiomyocytes with increasing concentrations of astemizole result in decreased peak frequency (middle panel) and arrhythmia (bottom panel) of the calcium transients after 30 minutes of incubation.

 

You can find the complete study in our application notes: