Abstract
Abstract A variety of physical and biochemical cell properties depends on the function and integrity of the actin cytoskeleton. The actin cytoskeleton mediates crucial cellular functions as migration, intracellular transport, exocytosis, endocytosis, cell stiffness and force generation. Highly dynamic actin fibers are therefore targets for several drugs and toxins. However, the study of actin interfering processes by standard microscopy techniques fails in the detailed resolution of dynamic spatial alterations required for a deeper understanding of toxic or drug effects. Here we applied digital holographic microscopy in the online functional analysis of the actin cytoskeleton changes of a highly differentiated and a dedifferentiated pancreas tumor cell line induced by the marine toxin Latrunculin B. Scanning electron microscopy (SEM) and fluorescence microscopy showed rapid Latrunculin B induced alterations in cell morphology and actin fiber degradation in both pancreatic tumor cell lines. In contrast digital holographic in vivo analysis of the drug dependent dynamic cellular processes revealed unequal changes in cell morphology. While tumor cells with a low metastatic potential showed Latrunculin B induced cell collapse within 4 h the metastatic tumor cells were resistant to Latrunculin B treatment. Spatial resolution of morphological alterations by digital holography detected so far unknown differences in the actin cytoskeleton stability of highly differentiated and dedifferentiated pancreas tumor cell lines. These data demonstrate that marker-free, non-destructive online analysis of cellular morphology and dynamic spatial processes in living cells by digital holography offers new insights in actin dependent cellular mechanisms. Digital holographic microscopy was shown to be a versatile tool in the screening of toxic drug effects and cancer cell biology.
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