The break-down and reassembly of myofibrils in long-term cultures of adult rat cardiomyocytes was investigated by a novel combination of confocal laser scanning microscopy and three-dimensional image reconstruction, referred to as FTCS, to visualize the morphological changes these cells undergo in culture. FTCS is discussed as an alternative imaging mode to low-magnification scanning electron microscopy. The three-dimensional shape of the cells are correlated with the assembly state of myofibrils in different stages. Based on immunofluorescence and confocal laser scanning microscopy it was shown that myofibrils are degraded within a few days after plating and that newly assembled myofibrils are predominantly confined to the continuous area in the perinuclear region close to the membrane in contact with the substratum. The localization of myofibrils along the cell's vertical axis has been investigated both by optical sectioning using confocal light microscopy and by physical sectioning followed by transmission electron microscopy. Based on the distribution of myofibrillar proteins we propose a model of myofibrillar growth locating the putative assembly sites to a region concentric around the nuclei. We provide evidence that the cell shape is dominated by the myofibrillar apparatus.
PVA coated and fluorescent dye (Cy3.5) functionalized vinyl alcohol/vinyl amine copolymer coated superparamagnetic iron oxide nanoparticles (SPION) were evaluated for systemic distribution and elimination after intraarticular injection in sheep. Observation was done at 3, 24, 72, and 120 hours after injection using light microscopy, fluorescent microscopy, and confocal microscopy. No pathologic influence of SPION on the tissue harvested could be seen. A significantly increased iron content could be identified in the kidneys, lymph nodes, and spleen after injection of SPION. No particles were detected in the liver, the urinary, and the gall bladder. No positive fluorescent signal could be attributed to SPION throughout the organs. Our results indicated that the iron component of the SPION is possible to be incorporated into the physiologic iron metabolism after reabsorption in the proximal tubule system of the kidney and that concentration levels of Cy3.5 are too low to be detected throughout the body.
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