Full volume super-resolution imaging of thick mitotic spindle using 3D AO STED microscope

Piotr Zdańkowski , David McGloin , Jason R. Swedlow

Abstract

Stimulated emission depletion (STED) nanoscopy is one of a suite of modern optical microscopy techniques capable of bypassing the conventional diffraction limit in fluorescent imaging. STED makes use of a spiral phase mask to enable 2D super-resolution imaging whereas to achieve full volumetric 3D super-resolution an additional bottle-beam phase mask must be applied. The resolution achieved in biological samples 10 µm or thicker is limited by aberrations induced mainly by scattering due to refractive index heterogeneity in the sample. These aberrations impact the fidelity of both types of phase mask, and have limited the application of STED to thicker biological systems. Here we apply an automated adaptive optics solution to correct the performance of both STED masks, enhancing robustness and expanding the capabilities of this nanoscopic technique. Corroboration in terms of successful high-quality imaging of the full volume of a 15µm mitotic spindle with resolution of 50nm x 50nm x 150nm achieved in all three dimensions is presented.
Author Piotr Zdańkowski (FM / IMPh)
Piotr Zdańkowski,,
- The Institute of Micromechanics and Photonics
, David McGloin - University of Dundee, Dundee, United Kingdom
David McGloin,,
-
, Jason R. Swedlow - University of Dundee, Dundee, United Kingdom
Jason R. Swedlow,,
-
Journal seriesBiomedical Optics Express, ISSN 2156-7085, (A 40 pkt)
Issue year2019
Vol10
No4
Pages1999-2009
Publication size in sheets0.5
Keywords in Englishmicroscopy, super-resolution microscopy, fluorescence microscopy, adaptive optics, aberration correction
ASJC Classification3107 Atomic and Molecular Physics, and Optics; 1305 Biotechnology
DOIDOI:10.1364/BOE.10.001999
URL https://www.osapublishing.org/boe/abstract.cfm?URI=boe-10-4-1999
Languageen angielski
Score (nominal)40
ScoreMinisterial score = 35.0, ArticleFromJournal
Ministerial score (2013-2016) = 40.0, ArticleFromJournal
Publication indicators Scopus SNIP (Source Normalised Impact per Paper): 2016 = 1.526; WoS Impact Factor: 2017 = 3.482 (2) - 2017=3.737 (5)
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