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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.

Original publication

DOI

10.1021/nl1027638

Type

Journal article

Journal

Nano Lett

Publication Date

13/10/2010

Volume

10

Pages

4249 - 4252

Keywords

Chlorophyll, Fluorescent Dyes, Microscopy, Fluorescence, Spinacia oleracea, Thylakoids