A New Wave of Cellular Imaging

  • Toomre D
  • Bewersdorf J
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Abstract

Fluorescence imaging methods that push or break the diffraction limit of
resolution (approximately 200 nm) have grown explosively. These
super-resolution nanoscopy techniques include: stimulated emission
depletion (STED), Pointillism microscopy {[}(fluorescence)
photoactivation localization microscopy/stochastic optical
reconstruction microscopy, or (F)PALM/STORM], structured illumination,
total internal reflection fluorescence microscopy (TIRFM), and those
that combine multiple modalities. Each affords unique strengths in
lateral and axial resolution, speed, sensitivity, and fluorophore
compatibility. We examine the optical principles and design of these new
instruments and their ability to see more detail with greater
sensitivity down to single molecules with tens of nanometers resolution.
Nanoscopes have revealed transient intermediate states of organelles and
molecules in living cells and have led to new discoveries but also
biological controversies. We highlight common unifying principles behind
nanoscopy such as the conversion of a subset of probes between states
(ground or excited) and the use of scanning (ordered or stochastic). We
emphasize major advances, biological applications, and promising new
developments.

Author-supplied keywords

  • super-resolution; light microscopy; total internal

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Authors

  • Derek Toomre

  • Joerg Bewersdorf

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