Concepts for structured illumination microscopy with extended axial resolution through mirrored illumination
Year: 2020
Type: article
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 I 5 M microscopy, have improved the axial resolution beyond that of the lateral, but at the cost of a significantly more complex optical setup. Here, we 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.
Source: Biomedical Optics Express
DOI: 10.1364/boe.382398
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,913
Funders: Cancer Prevention and Research Institute of Texas, Wellcome Trust, National Institutes of Health, Medical Research Council, Engineering and Physical Sciences Research Council, Fitzwilliam College, University of Cambridge, National Cancer Institute, National Institute of General Medical Sciences, H2020 Marie Skłodowska-Curie Actions
Awards: G1417656367, G1835608419, EPSRC Centre for Doctoral Training (CDT) in Sensor Technologies and Application, G2561383389, MICA: Multi-parametric and super-resolution imaging of amyloidogenic proteins, Development of single molecule techniques for nanoscale imaging of toxic protein species in vitro and in cells., G4454044550, Mechanisms of neurotoxicity of amyloid aggregates., Mechanisms of neurotoxicity of amyloid aggregates., Next generation live super-resolution microscopy: development and application at the Cambridge Advanced Imaging Centre, G5589109546, Development of Deep-UV Quantitative Microscopy for the Study of Mitochondrial Dysfunction, G5859390703, G6031874474, Recruitment of First-Time, Tenure-Track Faculty Members, G7413148874, Multiscale microscope for 3D cancer imaging in model organisms and organoids, Cambridge NanoScience through Engineering to Application Doctoral Training Centre: Assembly of Functional NanoMaterials and NanoDevices, G7823050896, G8039008064, Transformative microscopes to image across spatiotemporal scales
OpenAlex ID: W3006115974