Calcium flux in porcine left atrial appendage progenitor cells transfected with Premo™ Cameleon calcium sensor.
Porcine left atrial appendage progenitor cells were transfected with Premo™ Cameleon calcium sensor (P36207, P36208); ATP (20 µM final concentration) was applied to the cells the following day and the cells were imaged using a Zeiss 5 Live high-speed confocal system (Carl Zeiss MicroImaging). Excitation was with a 405 nm diode laser (50 mw) operated at 50% power. Emission was collected simultaneously on two linear CCD detectors using a 490 nm dichroic mirror to split the beam through a 415–480 nm bandpass filter for CFP and a 550 nm longpass filter for YFP. Images were collected at a rate of 10 frames per second (512 x 512 pixels) using a 40x Plan-Neofluar 1.3 NA oil immersion objective lens.