Open AccessComputational localization microscopy with extended axial range
Author(s) -
Yongzhuang Zhou,
Paul Zammit,
Guillem Carles,
Andrew R. Harvey
Publication year - 2018
Publication title -
optics express
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.26.007563
Subject(s) - optics , microscopy , materials science , image resolution , microscope , tracking (education) , optical tweezers , optical microscope , microfluidics , physics , scanning electron microscope , nanotechnology , psychology , pedagogy
A new single-aperture 3D particle-localization and tracking technique is presented that demonstrates an increase in depth range by more than an order of magnitude without compromising optical resolution and throughput. We exploit the extended depth range and depth-dependent translation of an Airy-beam PSF for 3D localization over an extended volume in a single snapshot. The technique is applicable to all bright-field and fluorescence modalities for particle localization and tracking, ranging from super-resolution microscopy through to the tracking of fluorescent beads and endogenous particles within cells. We demonstrate and validate its application to real-time 3D velocity imaging of fluid flow in capillaries using fluorescent tracer beads. An axial localization precision of 50 nm was obtained over a depth range of 120μm using a 0.4NA, 20× microscope objective. We believe this to be the highest ratio of axial range-to-precision reported to date.