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Nat Commun. 2016 Jul 25;7:12290. doi: 10.1038/ncomms12290.

Super-resolution spectroscopic microscopy via photon localization.

Author information

1
Biomedical Engineering Department, Northwestern University, Evanston, Illinois 60208, USA.
2
Mechanical Engineering Department, Northwestern University, Evanston, Illinois 60208, USA.
3
Advanced Imaging Center, Howard Hughes Medical Institute, Ashburn, Virginia 20147, USA.

Abstract

Traditional photon localization microscopy analyses only the spatial distributions of photons emitted by individual molecules to reconstruct super-resolution optical images. Unfortunately, however, the highly valuable spectroscopic information from these photons have been overlooked. Here we report a spectroscopic photon localization microscopy that is capable of capturing the inherent spectroscopic signatures of photons from individual stochastic radiation events. Spectroscopic photon localization microscopy achieved higher spatial resolution than traditional photon localization microscopy through spectral discrimination to identify the photons emitted from individual molecules. As a result, we resolved two fluorescent molecules, which were 15 nm apart, with the corresponding spatial resolution of 10 nm-a four-fold improvement over photon localization microscopy. Using spectroscopic photon localization microscopy, we further demonstrated simultaneous multi-colour super-resolution imaging of microtubules and mitochondria in COS-7 cells and showed that background autofluorescence can be identified through its distinct emission spectra.

PMID:
27452975
PMCID:
PMC4962472
DOI:
10.1038/ncomms12290
[Indexed for MEDLINE]
Free PMC Article

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