Far-field autofluorescence nanoscopy

Nano Lett. 2010 Oct 13;10(10):4249-52. doi: 10.1021/nl1027638.

Abstract

We demonstrate far-field optical imaging at the nanoscale with unlabeled samples. Subdiffraction resolution images of autofluorescent samples are obtained by depleting the ground state of natural fluorophores by transferring them to a metastable dark state and simultaneously localizing those fluorophores that are transiently returning. Our approach is based on the insight that nanoscopy methods relying on stochastic single-molecule switching require only a single fluorescence on-off cycle to yield an image, a condition fulfilled by various biomolecules. The method is exemplified by recording label-free nanoscopy images of thylakoid membranes of spinach chloroplasts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chlorophyll / analysis*
  • Fluorescent Dyes / analysis
  • Microscopy, Fluorescence / methods*
  • Spinacia oleracea / ultrastructure*
  • Thylakoids / ultrastructure*

Substances

  • Fluorescent Dyes
  • Chlorophyll