Sub-10-nm distance measurements between fluorophores using photon-accumulation enhanced reconstruction (PACER).

ABSTRACT: Single-molecule localization microscopy (SMLM) precisely localizes individual fluorescent molecules within the wide field of view. However, the localization precision is fundamentally limited to around 20 nm due to the physical photon limit of individual stochastic single-molecule emissions. Using spectroscopic SMLM (sSMLM) to resolve their distinct fluorescence emission spectra, we can specifically distinguish and identify individual fluorophore, even the ones of the same type. Consequently, the reported photon-accumulation enhanced reconstruction (PACER) method accumulates photons over repeated stochastic emissions from the same fluorophore to significantly improve the localization precision. This work showed the feasibility of PACER by resolving quantum dots that were 6.1 nm apart with

SUBMITTER: Dong B 

PROVIDER: S-EPMC7942786 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature