Deconvolution-free Subcellular Imaging with Axially Swept Light Sheet Microscopy

作者:Dean Kevin M; Roudot Philippe; Welf Erik S; Danuser Gaudenz; Fiolka Reto*
来源:Biophysical Journal, 2015, 108(12): 2807-2815.
DOI:10.1016/j.bpj.2015.05.013

摘要

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.

  • 出版日期2015-6-16