Concepts for structured illumination microscopy with extended axial resolution through mirrored illumination.
Biomedical optics express
Optical Society of America
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Manton, J., Ströhl, F., Fiolka, R., Kaminski, C., & Rees, E. J. (2020). Concepts for structured illumination microscopy with extended axial resolution through mirrored illumination.. Biomedical optics express, 11 (4), 2098-2108. https://doi.org/10.1364/boe.382398
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 a truly isotropic 3D resolution of 135 nm with conventional fluorophores and without the need for interferometric detection.
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)
Wellcome Trust (089703/Z/09/Z)
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External DOI: https://doi.org/10.1364/boe.382398
This record's URL: https://www.repository.cam.ac.uk/handle/1810/302688
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