The arrangement and movement of mitochondria were quantitatively studied in adult rat cardiomyocytes and in cultured continuously dividing non beating (NB) HL-1 cells with differentiated cardiac phenotype. Mitochondria were stained with MitoTracker Green and studied by fluorescent confocal microscopy. High speed scanning (one image every 400 ms) revealed very rapid fluctuation of positions of fluorescence centers of mitochondria in adult cardiomyocytes. These fluctuations followed the pattern of random walk movement within the limits of the internal space of mitochondria, probably due to transitions between condensed and orthodox configurational states of matrix and inner membrane. Mitochondrial fusion or fission was seen only in NB HL-1 cells but not in adult cardiomyocytes. In NB HL-1 cells, mitochondria were arranged as a dense tubular network, in permanent fusion, fission and high velocity displacements of approximately 90 nm/s. The differences observed in mitochondrial dynamics are related to specific structural organization and mitochondria-cytoskeleton interactions in these cells.