Single-beam multimodal nonlinear-optical imaging of structurally complex events in cell-cycle dynamics
A. А. Lanin, Artem S. Chebotarev, Ilya V. Kelmanson, M. S. Pochechuev, Elena S. Fetisova, Dmitry S. Bilan, Ekaterina Shevchenko, А. А. Иванов, A. B. Fedotov, Vsevolod V. Belousov, А. М. Желтиков
Abstract
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.