Fast-tracking of single emitters in large volumes with nanometer precision

Boris Louis, Rafael Camacho, Roger Bresoli-Obach, Sergey Abakumov, Johannes Vandaele, Testsuhiro Kudo, Hiroshi Masuhara, Ivan G. Scheblykin, Johan Hofkens, Susana Rocha

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Multifocal plane microscopy allows for capturing images at different focal planes simultaneously. Using a proprietary prism which splits the emitted light into paths of different lengths, images at 8 different focal depths were obtained, covering a volume of 50x50x4 mu m(3). The position of single emitters was retrieved using a phasor-based approach across the different imaging planes, with better than 10 nm precision in the axial direction. We validated the accuracy of this approach by tracking fluorescent beads in 3D to calculate water viscosity. The fast acquisition rate (>100 fps) also enabled us to follow the capturing of 0.2 mu m fluorescent beads into an optical trap. (C) 2020 Optical Society of America under the terms of the OSA Open Access Publishing Agreement

Original languageEnglish
Article number401557
Pages (from-to)28656-28671
Number of pages16
JournalOptics Express
Volume28
Issue number19
DOIs
StatePublished - 14 Sep 2020

Keywords

  • 3-DIMENSIONAL PARTICLE TRACKING
  • IMAGE CORRELATION SPECTROSCOPY
  • MULTIFOCAL PLANE MICROSCOPY
  • FLUORESCENCE MICROSCOPY
  • LOCALIZATION MICROSCOPY
  • MOLECULE LOCALIZATION
  • CELL
  • DYNAMICS
  • DIFFUSION
  • RECEPTOR

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