中央研究院 原子與分子科學研究所

Nanoparticles have been used extensively in biology-related research and many applications require direct visualization of individual nanoparticles under optical microscopy. For long-term and high-speed measurements, scattering-based microscopy is a unique technique because of the stable and indefinite scattering signal. In scattering-based single-particle measurements, large nanoparticles are usually needed in order to generate sufficient signal for detection. However, larger nanoparticle introduces greater mass loading, experiences stronger steric hindrance, and is more prone to crosslinking. In this work, we demonstrate coherent brightfield (COBRI) microscopy with enhanced contrast and show its capability of direct visualization of very small nanoparticles in scattering at high speed. The COBRI microscopy allows us to visualize and track single metallic and dielectric nanoparticles, as small as 10 nm, at 1,000 frames per second. A quantitative relationship between the linear scattering cross section of nanoparticle and its COBRI contrast is reported. Using COBRI microscopy, we further demonstrate tracking of 10 nm gold nanoparticles labeled to lipid molecules in supported bilayer membranes, showing that the small nanoparticle may facilitate single-molecule measurements with reduced perturbation. Furthermore, identical imaging sensitivity of COBRI and interferometric scattering (iSCAT) microscopy, the reflection counterpart of COBRI, is demonstrated under equal illumination intensity. Finally, future improvements in speed and sensitivity of scattering-based interference microscope are discussed.

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