Three-Dimensional Unstained Live-Cell Imaging Using Stimulated Parametric Emission Microscopy

Abstract

The ability to perform high-resolution unstained live imaging is very important to in vivo study of cell structures and functions. Stimulated parametric emission (SPE) microscopy is a nonlinear-optical microscopy based on ultra-fast electronic nonlinear-optical responses. For the first time, we have successfully applied this technique to archive three-dimensional (3D) images of unstained sub-cellular structures, such as, microtubules, nuclei, nucleoli, etc. in live cells. Observation of a complete cell division confirms the ability of SPE microscopy for long time-scale imaging.

Visualization of Oil Body Distribution in Jatropha curcas L. by Four-Wave Mixing Microscopy

“…These advantages of SPE microscopy have also been reported for other biological samples. 24) Thus, we have successfully applied SPE microscopy to the imaging of oil bodies in jatropha kernels without staining or labeling.…”

“…Mechanism (i) was demonstrated by the SPE imaging of unstained blood cells, 22) a stained mouse kidney tissue, and an unstained purple petal tissue of Petunia hybrida. 23) Mechanism (ii) was demonstrated by the visualization of subcellular structures of an unstained living cell, 24) in addition to RI profiling. 25) Here, we demonstrate that SPE microscopy based on mechanism (ii) can be employed to clearly visualize oil distribution of a jatropha kernel without staining or labeling.…”