dc.contributor.author | Coucheron, David Andre | |
dc.contributor.author | Helle, Øystein Ivar | |
dc.contributor.author | Øie, Cristina Ionica | |
dc.contributor.author | Tinguely, Jean-Claude | |
dc.contributor.author | Ahluwalia, Balpreet Singh | |
dc.date.accessioned | 2020-02-19T10:59:01Z | |
dc.date.available | 2020-02-19T10:59:01Z | |
dc.date.issued | 2019-11-16 | |
dc.description.abstract | Total internal reflection fluorescence (TIRF) is commonly used in single molecule localization based super-resolution microscopy as it gives enhanced contrast due to optical sectioning. The conventional approach is to use high numerical aperture microscope TIRF objectives for both excitation and collection, severely limiting the field of view and throughput. We present a novel approach to generating TIRF excitation for imaging with optical waveguides, called chip-based nanoscopy. The aim of this protocol is to demonstrate how chip-based imaging is performed in an already built setup. The main advantage of chip-based nanoscopy is that the excitation and collection pathways are decoupled. Imaging can then be done with a low magnification lens, resulting in large field of view TIRF images, at the price of a small reduction in resolution. Liver sinusoidal endothelial cells (LSECs) were imaged using <i>direct</i> stochastic optical reconstruction microscopy (<i>d</i>STORM), showing a resolution comparable to traditional super-resolution microscopes. In addition, we demonstrate the high-throughput capabilities by imaging a 500 µm x 500 µm region with a low magnification lens, providing a resolution of 76 nm. Through its compact character, chip-based imaging can be retrofitted into most common microscopes and can be combined with other on-chip optical techniques, such as on-chip sensing, spectroscopy, optical trapping, etc. The technique is thus ideally suited for high throughput 2D super-resolution imaging, but also offers great opportunities for multi-modal analysis. | en_US |
dc.identifier.citation | Coucheron DA, Helle ØI, Øie CI, Tinguely J, Ahluwalia BS. High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip. Journal of Visualized Experiments. 2019 | en_US |
dc.identifier.cristinID | FRIDAID 1749300 | |
dc.identifier.doi | http://dx.doi.org/10.3791/60378 | |
dc.identifier.issn | 1940-087X | |
dc.identifier.uri | https://hdl.handle.net/10037/17423 | |
dc.language.iso | eng | en_US |
dc.publisher | Journal of Visualized Experiments | en_US |
dc.relation.ispartof | Coucheron, D.A. (2021). Waveguide-based Excitation for High-throughput Imaging. (Doctoral thesis). <a href=https://hdl.handle.net/10037/20695>https://hdl.handle.net/10037/20695</a> | |
dc.relation.journal | Journal of Visualized Experiments | |
dc.relation.projectID | info:eu-repo/grantAgreement/EC/FP7-IDEAS-ERC/336716/EU/High-speed chip-based nanoscopy to discover real-time sub-cellular dynamics/NANOSCOPY/ | |
dc.rights.accessRights | openAccess | en_US |
dc.rights.holder | Copyright 2019 The Author(s) | en_US |
dc.subject | VDP::Mathematics and natural science: 400 | en_US |
dc.subject | VDP::Matematikk og Naturvitenskap: 400 | en_US |
dc.title | High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip | en_US |
dc.type.version | publishedVersion | en_US |
dc.type | Journal article | en_US |
dc.type | Tidsskriftartikkel | en_US |
dc.type | Peer reviewed | en_US |