High-speed microscopy with an electrically tunable lens to image the dynamics of in vivo molecular complexes

• DOI: 10.1063/1.4905330
• Publication Date: 2015-01-20T18:17:47Z
• Authors: Yuichiro Nakai; Mitsunori Ozeki; Takumi Hiraiwa; Ryuichi Tanimoto; Akira Funahashi; Noriko Hiroi; Atsushi Taniguchi; Shigenori Nonaka; Viviane Boilot; Roshan Shrestha; Joanna Clark; Naoka Tamura; Viji M. Draviam; Hiromasa Oku
• Keywords: 0301 basic medicine; Electrical Equipment and Supplies; Green Fluorescent Proteins; Intracellular Space; Video Recording; Proteins; Microtubules; Pattern Recognition, Automated; 03 medical and health sciences; Imaging, Three-Dimensional; Microscopy, Fluorescence; Humans; Computer Simulation; HeLa Cells; Lenses

We provide an evaluation for an electrically tunable lens (ETL), combined with a microscope system, from the viewpoint of tracking intracellular protein complexes. We measured the correlation between the quantitative axial focus shift and the control current for ETL, and determined the stabilization time for refocusing to evaluate the electrical focusing behaviour of our system. We also confirmed that the change of relative magnification by the lens and associated resolution does not influence the ability to find intracellular targets. By applying the ETL system to observe intracellular structures and protein complexes, we confirmed that this system can obtain 10 nm order z-stacks, within video rate, while maintaining the quality of images and that this system has sufficient optical performance to detect the molecules.