Pulsed Interleaved MINFLUX

We introduce p-MINFLUX, a new implementation of the highly photon-efficient single-molecule localization method with a simplified experimental setup and additional fluorescence lifetime information. In contrast to the original MINFLUX implementation, p-MINFLUX uses interleaved laser pulses to delive...

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Bibliographic Details
Published in:Nano letters 2021-01, Vol.21 (1), p.840-846
Main Authors: Masullo, Luciano A, Steiner, Florian, Zähringer, Jonas, Lopez, Lucía F, Bohlen, Johann, Richter, Lars, Cole, Fiona, Tinnefeld, Philip, Stefani, Fernando D
Format: Article
Language:English
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Summary:We introduce p-MINFLUX, a new implementation of the highly photon-efficient single-molecule localization method with a simplified experimental setup and additional fluorescence lifetime information. In contrast to the original MINFLUX implementation, p-MINFLUX uses interleaved laser pulses to deliver the doughnut-shaped excitation foci at a maximum repetition rate. Using both static and dynamic DNA origami model systems, we demonstrate the performance of p-MINFLUX for single-molecule localization nanoscopy and tracking, respectively. p-MINFLUX delivers 1-2 nm localization precision with 2000-1000 photon counts. In addition, p-MINFLUX gives access to the fluorescence lifetime enabling multiplexing and super-resolved lifetime imaging. p-MINFLUX should help to unlock the full potential of innovative single-molecule localization schemes.
ISSN:1530-6984
1530-6992
DOI:10.1021/acs.nanolett.0c04600