Abstract
Dynamic imaging of cardiomyocytes was performed with a simultaneous detection second harmonic generation (SHG), third harmonic generation (THG) and multiphoton excitation fluorescence (MPF) microscope. The fast scanning system of ~12 frames/second synchronized with multichannel detection provided the possibility of imaging three dimensional static and two dimensional dynamic structures of cardiomyocytes. The SHG images highlighted the myofibrils of the cardiomyocytes while THG images revealed the locations of mitochondria. Dynamic data showed that during imaging, chaotic nanocontractions took place inside the cardiomyocytes. The time series of THG images reveled large intensity fluctuations "flickering" in the regions of mitochondria. The flickering in THG correlated with the flickering in MPF. Addition of the uncoupler FCCP inhibited flickering in THG and MPF, and also inhibited nanocontractions. The simultaneous imaging with SHG, THG and MPF proved to be a very powerful microscopy tool for investigation of interactions of different organelles inside a cell.
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