Deconvolution-free Subcellular Imaging with Axially Swept Light Sheet Microscopy.

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.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Dean KM,Roudot P,Welf ES,Danuser G,Fiolka R

doi

10.1016/j.bpj.2015.05.013

subject

Has Abstract

pub_date

2015-06-16 00:00:00

pages

2807-15

issue

12

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(15)00498-1

journal_volume

108

pub_type

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