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

Rev Sci Instrum. 2015 Jan;86(1):013707. doi: 10.1063/1.4905330.

Abstract

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.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Computer Simulation
  • Electrical Equipment and Supplies*
  • Green Fluorescent Proteins / metabolism
  • HeLa Cells
  • Humans
  • Imaging, Three-Dimensional / instrumentation
  • Imaging, Three-Dimensional / methods
  • Intracellular Space / metabolism*
  • Lenses*
  • Microscopy, Fluorescence / instrumentation*
  • Microscopy, Fluorescence / methods*
  • Microtubules / metabolism
  • Pattern Recognition, Automated
  • Proteins / metabolism*
  • Video Recording / instrumentation
  • Video Recording / methods

Substances

  • Proteins
  • Green Fluorescent Proteins