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

LOUIS, BORIS LU ; CAMACHO, RAFAEL ; BRESOLÍ-OBACH, ROGER ; ABAKUMOV, SERGEY ; VANDAELE, JOHANNES ; KUDO, TESTSUHIRO ; MASUHARA, HIROSHI ; SCHEBLYKIN, IVAN G. LU orcid ; HOFKENS, JOHAN and ROCHA, SUSANA (2020) In Optics Express 28(19). p.28656-28671
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 μm3. 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 μm fluorescent beads into an optical trap.

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author
; ; ; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Optics Express
volume
28
issue
19
article number
401557
pages
16 pages
publisher
Optical Society of America
external identifiers
  • scopus:85091845720
  • pmid:32988132
ISSN
1094-4087
DOI
10.1364/OE.401557
language
English
LU publication?
yes
id
a1c27d26-2d61-4d03-a9ec-a74f12d9e756
date added to LUP
2020-11-02 13:22:59
date last changed
2024-05-15 19:56:05
@article{a1c27d26-2d61-4d03-a9ec-a74f12d9e756,
  abstract     = {{<p>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 μm3. 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 (&gt;100 fps) also enabled us to follow the capturing of 0.2 μm fluorescent beads into an optical trap.</p>}},
  author       = {{LOUIS, BORIS and CAMACHO, RAFAEL and BRESOLÍ-OBACH, ROGER and ABAKUMOV, SERGEY and VANDAELE, JOHANNES and KUDO, TESTSUHIRO and MASUHARA, HIROSHI and SCHEBLYKIN, IVAN G. and HOFKENS, JOHAN and ROCHA, SUSANA}},
  issn         = {{1094-4087}},
  language     = {{eng}},
  number       = {{19}},
  pages        = {{28656--28671}},
  publisher    = {{Optical Society of America}},
  series       = {{Optics Express}},
  title        = {{Fast-tracking of single emitters in large volumes with nanometer precision}},
  url          = {{http://dx.doi.org/10.1364/OE.401557}},
  doi          = {{10.1364/OE.401557}},
  volume       = {{28}},
  year         = {{2020}},
}