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A transparent waveguide chip for versatile total internal reflection fluorescence-based microscopy and nanoscopy

A transparent waveguide chip for versatile total internal reflection fluorescence-based microscopy and nanoscopy
A transparent waveguide chip for versatile total internal reflection fluorescence-based microscopy and nanoscopy
Total internal reflection fluorescence (TIRF) microscopy is an imaging technique that, in comparison to confocal microscopy, does not require a trade-off between resolution, speed, and photodamage. Here, we introduce a waveguide platform for chip-based TIRF imaging based on a transparent substrate, which is fully compatible with sample handling and imaging procedures commonly used with a standard #1.5 glass coverslip. The platform is fabricated using standard complementary metal-oxide-semiconductor techniques which can easily be scaled up for mass production. We demonstrate its performance on synthetic and biological samples using both upright and inverted microscopes, and show how it can be extended to super-resolution applications, achieving a resolution of 116 nm using super resolution radial fluctuations. These transparent chips retain the scalable field of view of opaque chip-based TIRF and the high axial resolution of TIRF, and have the versatility to be used with many different objective lenses, microscopy methods, and handling techniques. We see this as a technology primed for widespread adoption, increasing both TIRF’s accessibility to users and the range of applications that can benefit from it.
2662-4443
Priyadarshi, Anish
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Dullo, Firehun
4a6bb735-5a04-4283-92e5-9506d46f6b18
Wolfson, Deanna Lynn
2d0925fb-41e1-4071-a105-cc9337857d42
Ahmad, Azeem
c8d1e19a-904a-43ae-a9a4-2b35b8670f62
Jayakumar, Nikhil
74426c48-7dc2-4181-b258-0dcd5dc7d28a
Dubey, Vishesh
778bd79c-07e9-4ca3-ad2e-8aead242d081
Tinguely, Jean-Claude
b5a06cdf-d7b3-474b-9c4f-f3e4926e2db8
Ahluwalia, Balpreet
1a4a978b-ae2b-478d-ac6b-126c08361a65
Murugan, Ganapathy Senthil
a867686e-0535-46cc-ad85-c2342086b25b
Priyadarshi, Anish
917c518c-adce-4f07-a88e-4716c2f83dc9
Dullo, Firehun
4a6bb735-5a04-4283-92e5-9506d46f6b18
Wolfson, Deanna Lynn
2d0925fb-41e1-4071-a105-cc9337857d42
Ahmad, Azeem
c8d1e19a-904a-43ae-a9a4-2b35b8670f62
Jayakumar, Nikhil
74426c48-7dc2-4181-b258-0dcd5dc7d28a
Dubey, Vishesh
778bd79c-07e9-4ca3-ad2e-8aead242d081
Tinguely, Jean-Claude
b5a06cdf-d7b3-474b-9c4f-f3e4926e2db8
Ahluwalia, Balpreet
1a4a978b-ae2b-478d-ac6b-126c08361a65
Murugan, Ganapathy Senthil
a867686e-0535-46cc-ad85-c2342086b25b

Priyadarshi, Anish, Dullo, Firehun, Wolfson, Deanna Lynn, Ahmad, Azeem, Jayakumar, Nikhil, Dubey, Vishesh, Tinguely, Jean-Claude, Ahluwalia, Balpreet and Murugan, Ganapathy Senthil (2021) A transparent waveguide chip for versatile total internal reflection fluorescence-based microscopy and nanoscopy. Communications Materials, 2 (1), [85]. (doi:10.1038/s43246-021-00192-5).

Record type: Article

Abstract

Total internal reflection fluorescence (TIRF) microscopy is an imaging technique that, in comparison to confocal microscopy, does not require a trade-off between resolution, speed, and photodamage. Here, we introduce a waveguide platform for chip-based TIRF imaging based on a transparent substrate, which is fully compatible with sample handling and imaging procedures commonly used with a standard #1.5 glass coverslip. The platform is fabricated using standard complementary metal-oxide-semiconductor techniques which can easily be scaled up for mass production. We demonstrate its performance on synthetic and biological samples using both upright and inverted microscopes, and show how it can be extended to super-resolution applications, achieving a resolution of 116 nm using super resolution radial fluctuations. These transparent chips retain the scalable field of view of opaque chip-based TIRF and the high axial resolution of TIRF, and have the versatility to be used with many different objective lenses, microscopy methods, and handling techniques. We see this as a technology primed for widespread adoption, increasing both TIRF’s accessibility to users and the range of applications that can benefit from it.

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e-pub ahead of print date: 20 August 2021
Published date: 20 August 2021

Identifiers

Local EPrints ID: 451066
URI: http://eprints.soton.ac.uk/id/eprint/451066
ISSN: 2662-4443
PURE UUID: 2bddab5a-73b7-4771-900a-16b54a3bf655
ORCID for Ganapathy Senthil Murugan: ORCID iD orcid.org/0000-0002-2733-3273

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Date deposited: 06 Sep 2021 16:31
Last modified: 17 Mar 2024 03:03

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Contributors

Author: Anish Priyadarshi
Author: Firehun Dullo
Author: Deanna Lynn Wolfson
Author: Azeem Ahmad
Author: Nikhil Jayakumar
Author: Vishesh Dubey
Author: Jean-Claude Tinguely
Author: Balpreet Ahluwalia

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