Pulsed Interleaved MINFLUX

MASULLO, LUCIANO A.; STEINER, FLORIAN; ZÄHRINGER, JONAS; LOPEZ, LUCÍA F.; BOHLEN, JOHANN; RICHTER, LARS; COLE, FIONA; TINNEFELD, PHILIP; STEFANI, FERNANDO D.

NANO LETTERS (PRINT)

AMER CHEMICAL SOC

Año: 2020 vol. 21 p. 840 - 846

1530-6984

We introduce p-MINFLUX, a new implementation ofthe highly photon-efficient single-molecule localization method witha simplified experimental setup and additional fluorescence lifetimeinformation. In contrast to the original MINFLUX implementation,p-MINFLUX uses interleaved laser pulses to deliver the doughnutshapedexcitation foci at a maximum repetition rate. Using both staticand dynamic DNA origami model systems, we demonstrate theperformance of p-MINFLUX for single-molecule localization nanoscopyand tracking, respectively. p-MINFLUX delivers 1−2 nmlocalization precision with 2000−1000 photon counts. In addition, p-MINFLUX gives access to the fluorescence lifetime enablingmultiplexing and super-resolved lifetime imaging. p-MINFLUXshould help to unlock the full potential of innovative single-moleculelocalization schemes.