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Concepts for structured illumination microscopy with extended axial resolution through mirrored illumination.

Accepted version
Peer-reviewed

Type

Article

Change log

Authors

Fiolka, Reto 
Kaminski, Clemens F 
Rees, Eric J 

Abstract

Wide-field fluorescence microscopy, while much faster than confocal microscopy, suffers from a lack of optical sectioning and poor axial resolution. 3D structured illumination microscopy (SIM) has been demonstrated to provide optical sectioning and to double the resolution limit both laterally and axially, but even with this the axial resolution is still worse than the lateral resolution of unmodified wide-field microscopy. Interferometric schemes using two high numerical aperture objectives, such as 4Pi confocal and I5M microscopy, have improved the axial resolution beyond that of the lateral, but at the cost of a significantly more complex optical setup. Here, we theoretically and numerically investigate a simpler dual-objective scheme which we propose can be easily added to an existing 3D-SIM microscope, providing lateral and axial resolutions in excess of 125 nm with conventional fluorophores and without the need for interferometric detection.

Description

Keywords

51 Physical Sciences, 40 Engineering, 4018 Nanotechnology, Bioengineering, 1 Underpinning research, 1.5 Resources and infrastructure (underpinning)

Journal Title

Biomed Opt Express

Conference Name

Journal ISSN

2156-7085
2156-7085

Volume Title

11

Publisher

Optica Publishing Group

Rights

All rights reserved
Sponsorship
EPSRC (1503810)
Engineering and Physical Sciences Research Council (EP/H018301/1)
Engineering and Physical Sciences Research Council (EP/G037221/1)
Wellcome Trust (089703/Z/09/Z)
Medical Research Council (MR/K015850/1)
Medical Research Council (MR/K02292X/1)
Engineering and Physical Sciences Research Council (EP/L015889/1)
CFK acknowledges funding from the UK EPSRC (EP/L015889/1, EP/H018301/1 and EP/G037221/1), the Wellcome Trust (3-3249/Z/16/Z and 089703/Z/09/Z), the UK MRC (MR/K015850/1 and MR/K02292X/1)
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