Deconvolution-free Subcellular Imaging with Axially Swept Light Sheet Microscopy

Kevin M. Dean, Philippe Roudot, Erik S. Welf, Gaudenz Danuser, Reto Fiolka

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

The use of propagation invariant Bessel beams has enabled high-resolution subcellular light sheet fluorescence microscopy. However, the energy within the concentric side lobe structure of Bessel beams increases significantly with propagation length, generating unwanted out-of-focus fluorescence that enforces practical limits on the imaging field of view size. Here, we present a light sheet fluorescence microscope that achieves 390 nm isotropic resolution and high optical sectioning strength (i.e., out-of-focus blur is strongly suppressed) over large field of views, without the need for structured illumination or deconvolution-based postprocessing. We demonstrate simultaneous dual-color, high-contrast, and high-dynamic-range time-lapse imaging of migrating cells in complex three-dimensional microenvironments, three-dimensional tracking of clathrin-coated pits, and long-term imaging spanning >10 h and encompassing >2600 time points.

Original languageEnglish (US)
Pages (from-to)2807-2815
Number of pages9
JournalBiophysical Journal
Volume108
Issue number12
DOIs
StatePublished - Jun 18 2015

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Microscopy
Fluorescence
Time-Lapse Imaging
Light
Clathrin
Lighting
Fluorescence Microscopy
Color

ASJC Scopus subject areas

  • Biophysics

Cite this

Deconvolution-free Subcellular Imaging with Axially Swept Light Sheet Microscopy. / Dean, Kevin M.; Roudot, Philippe; Welf, Erik S.; Danuser, Gaudenz; Fiolka, Reto.

In: Biophysical Journal, Vol. 108, No. 12, 18.06.2015, p. 2807-2815.

Research output: Contribution to journalArticle

Dean, Kevin M. ; Roudot, Philippe ; Welf, Erik S. ; Danuser, Gaudenz ; Fiolka, Reto. / Deconvolution-free Subcellular Imaging with Axially Swept Light Sheet Microscopy. In: Biophysical Journal. 2015 ; Vol. 108, No. 12. pp. 2807-2815.
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