3-DIMENSIONAL ANALYSIS AND VISUALIZATION OF MYOFIBRILLOGENESIS IN ADULT CARDIOMYOCYTES BY CONFOCAL MICROSCOPY

Confocal light microscopy has found its place among the standard analy tical tools in cell and molecular biology. When combined with techniqu es such as immunofluorescence or fluorescent in situ hybridization, th e spatial distribution of individual biological components can be trac ed within cells and tissues and, under certain circumstances, even wit h living samples. In this article, advanced 3D visualization technique s have been applied to analyze the distribution of myofibrillar protei ns in cultured adult rat cardiomyocytes. By combining confocal immunof luorescence microscopy with specially designed three-dimensional visua lization, we have obtained images which are similar to those obtained with the scanning electron microscope. The subcellular distribution of proteins expressed after transfection of cDNA is monitored in the cul tured heart cells. The expressed proteins are distinguished from their endogenous counterparts by the use of an epitope tagging technique. T he described methods are suitable to specifically monitor the behavior of several closely related isoprotein mutants in cell or tissue prepa rations. (C) 1995 Wiley-Liss, Inc.