Single-beam multimodal nonlinear-optical imaging of structurally complex events in cell-cycle dynamics

Abstract

We demonstrate a multimodal nonlinear-optical imaging that combines second- and third-harmonic generation (SHG and THG) with three-photon-excited fluorescence (3PEF) as a means to resolve fine details of the cell structure and trace its transformations throughout structurally complex episodes of cell-cycle dynamics, including the key stages and signatures in cell division. When zoomed in on cell mitosis, this technique enables a high-contrast multimodal imaging of intra- and extracellular signatures of cell division, detecting, via a multiplex, 3PEF/SHG/THG readout, a remarkable diversity of shapes, sizes, and symmetries in a truly single-beam setting, with no need for beam refocusing or field-waveform re-adjustment.