Curd, AP, Cleasby, A, Makowska, K et al. (3 more authors) (2015) Construction of an instant structured illumination microscope. Methods, 88. 37 - 47. ISSN 1046-2023
Abstract
A challenge in biological imaging is to capture high-resolution images at fast frame rates in live cells. The “instant structured illumination microscope” (iSIM) is a system designed for this purpose. Similarly to standard structured illumination microscopy (SIM), an iSIM provides a twofold improvement over widefield microscopy, in x, y and z, but also allows much faster image acquisition, with real-time display of super-resolution images. The assembly of an iSIM is reasonably complex, involving the combination and alignment of many optical components, including three micro-optics arrays (two lenslet arrays and an array of pinholes, all with a pitch of 222 μm) and a double-sided scanning mirror. In addition, a number of electronic components must be correctly controlled. Construction of the system is therefore not trivial, but is highly desirable, particularly for live-cell imaging. We report, and provide instructions for, the construction of an iSIM, including minor modifications to a previous design in both hardware and software. The final instrument allows us to rapidly acquire fluorescence images at rates faster than 100 fps, with approximately twofold improvement in resolution in both x-y and z; sub-diffractive biological features have an apparent size (full width at half maximum) of 145 nm (lateral) and 320 nm (axial), using a 1.49 NA objective and 488 nm excitation.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2015, The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
Keywords: | Construction; Fluorescence microscopy; Instant structured illumination microscope; Super-resolution |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) |
Funding Information: | Funder Grant number MRC MR/K015613/1 |
Depositing User: | Symplectic Publications |
Date Deposited: | 30 Jul 2015 13:04 |
Last Modified: | 26 Feb 2019 09:36 |
Published Version: | http://dx.doi.org/10.1016/j.ymeth.2015.07.012 |
Status: | Published |
Publisher: | Elsevier |
Identification Number: | 10.1016/j.ymeth.2015.07.012 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:88610 |
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