A hybrid high-speed atomic force-optical microscope for visualizing single membrane proteins on eukaryotic cells. Academic Article uri icon

Overview

abstract

  • High-speed atomic force microscopy is a powerful tool for studying structure and dynamics of proteins. So far, however, high-speed atomic force microscopy was restricted to well-controlled molecular systems of purified proteins. Here we integrate an optical microscopy path into high-speed atomic force microscopy, allowing bright field and fluorescence microscopy, without loss of high-speed atomic force microscopy performance. This hybrid high-speed atomic force microscopy/optical microscopy setup allows positioning of the high-speed atomic force microscopy tip with high spatial precision on an optically identified zone of interest on cells. We present movies at 960 ms per frame displaying aquaporin-0 array and single molecule dynamics in the plasma membrane of intact eye lens cells. This hybrid setup allows high-speed atomic force microscopy imaging on cells about 1,000 times faster than conventional atomic force microscopy/optical microscopy setups, and allows first time visualization of unlabelled membrane proteins on a eukaryotic cell under physiological conditions. This development advances high-speed atomic force microscopy from molecular to cell biology to analyse cellular processes at the membrane such as signalling, infection, transport and diffusion.

publication date

  • January 1, 2013

Research

keywords

  • Aquaporins
  • Cell Membrane
  • Escherichia coli
  • Eye Proteins
  • Lens, Crystalline
  • Microscopy, Atomic Force
  • Microscopy, Fluorescence

Identity

Scopus Document Identifier

  • 84880364773

Digital Object Identifier (DOI)

  • 10.1038/ncomms3155

PubMed ID

  • 23857417

Additional Document Info

volume

  • 4