Being a label-free technique, iSCAT has emerged as a powerful tool complementary to fluorescence microscopy because its superb spatio-temporal resolution holds great promise in biological investigations ranging from detection of nm-sized proteins to real-time dynamics of organelles in vivo. Here we present the polarization-selective iSCAT (psiSCAT) microscopy to detect the orientation and anisotropy of nano-objects. We also succeeded in developing the remote-focusing iSCAT (RF-iSCAT) to realize fast, vibration-free, 3D imaging over a long z-tracking range (< 5 μm). We envisage that our achievement would greatly advance the field of single-particle tracking into new dimensions with superb precision and sensitivity.
We introduce the cargo-localization iSCAT microscopy, label-free, live-cell imaging technique, that visualizes the traffic of cargos in a massively parallel fashion and hence uncovers the actively-used cytoskeletal network from cargos’ traces. By applying a particle localization algorithm to over a hundred cargos moving in a lamellipodium, the fine architecture of the cytoskeletal highway could be reconstructed. We also discuss a host of interesting kinetic events associated with cargo transport such as intermittent pausing, turnover, and frequent jamming of cargos due to physical collision with other obstacles or at the intersection of the highway.
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