Monitoring ruffling cells by lattice light-sheet microscopy

摘要

Membrane ruffling is an essential process at the leading edge of the migrating cells, which contains protrusion and retraction of plasma membrane. The extension membrane determines the direction of migration. The dynamic of membrane ruffling depends on the interaction between filament actin and motor proteins. Upon the activation of motor proteins by calcium ions, the migration process starts. Therefore, it is important to study the correlation between local calcium concentration and the membrane dynamics. To study the dynamics of the membrane ruffling, we established several stable cell lines, which contain chemical and optogenetic inducible dimerization . In addition, the proteins of interest such as actin, myosin, membrane anchors and calcium indicators were labeled with fluorescence proteins. The dynamics of membrane ruffling was investigated by lattice light-sheet microscope (LLSM), which is capable of high spatial and temporal recording over three-dimensions.