Deconvolution-free Subcellular Imaging with Axially Swept Light Sheet Microscopy

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...

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Detalles Bibliográficos
Autores principales: Dean, Kevin M., Roudot, Philippe, Welf, Erik S., Danuser, Gaudenz, Fiolka, Reto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Biophysical Society 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4472079/
https://www.ncbi.nlm.nih.gov/pubmed/26083920
http://dx.doi.org/10.1016/j.bpj.2015.05.013
Descripción
Sumario: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.