Far-field autofluorescence nanoscopy.
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.
Bierwagen
Jakob
J
Max Planck Institute for Biophysical Chemistry, Department of NanoBiophotonics, Am Fassberg 11, 37077 Gttingen, Germany.
Testa
Ilaria
I
Fölling
Jonas
J
Wenzel
Dirk
D
Jakobs
Stefan
S
Eggeling
Christian
C
Hell
Stefan W
SW
D002734:Chlorophyll / Q000032:analysis*; D005456:Fluorescent Dyes / Q000032:analysis; D008856:Microscopy, Fluorescence / Q000379:methods*; D018724:Spinacia oleracea / Q000648:ultrastructure*; D020524:Thylakoids / Q000648:ultrastructure*
D005456:Fluorescent Dyes; D002734:Chlorophyll