Unlike those of other ordinary laser scanning microscopies in the past, nonlinear optical laser scanning microscopy (SHG, THG microscopy) applied ultrafast laser technology which has high peak powers with relatively inexpensive, low-average-power. It short pulse nature reduces the ionization damage in organic molecules. And it enables us to take bright label-free images. In this study, we measured cell division of zebrafish egg with ultrafast video images using multimodal nonlinear optical microscope. The result shows in-vivo cell division label-free imaging with sub-cellular resolution.
Nonlinear optical microscopy has enabled the possibility to explore inside the living organisms. It utilizes ultrashort laser pulse with long wavelength (greater than 800nm). Ultrashort pulse produces high peak power to induce nonlinear optical phenomenon such as two-photon excitation fluorescence (TPEF) and harmonic generations in the medium while maintaining relatively low average energy pre area. In plant developmental biology, confocal microscopy is widely used in plant cell imaging after the development of biological fluorescence labels in mid-1990s. However, fluorescence labeling itself affects the sample and the sample deviates from intact condition especially when labelling the entire cell. In this work, we report the dynamic images of Arabidopsis thaliana root cells. This demonstrates the multimodal nonlinear optical microscopy is an effective tool for long-term plant cell imaging.