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Nanoscopy with more than 100,000 'doughnuts'.

Andriy ChmyrovJan KellerTim GrotjohannMichael RatzElisa d'EsteStefan JakobsChristian EggelingStefan W Hell
Published in: Nature methods (2013)
We show that nanoscopy based on the principle called RESOLFT (reversible saturable optical fluorescence transitions) or nonlinear structured illumination can be effectively parallelized using two incoherently superimposed orthogonal standing light waves. The intensity minima of the resulting pattern act as 'doughnuts', providing isotropic resolution in the focal plane and making pattern rotation redundant. We super-resolved living cells in 120 μm × 100 μm-sized fields of view in <1 s using 116,000 such doughnuts.
Keyphrases
  • living cells
  • single molecule
  • fluorescent probe
  • high intensity
  • high speed
  • energy transfer