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New super-oscillatory technology for unlabelled super-resolution cellular imaging with polarisation contrast

New super-oscillatory technology for unlabelled super-resolution cellular imaging with polarisation contrast
New super-oscillatory technology for unlabelled super-resolution cellular imaging with polarisation contrast
Super-resolution microscopy is already showing huge benefits across the biosciences, but all widely-used techniques require the addition of fluorescent probes. We have demonstrated optical-super-resolution imaging in unlabelled living cells, using the phenomenon of super-oscillation.

Super-oscillation is originally a mathematical phenomenon, first described in quantum mechanics. It is widely accepted that any function that is band-limited (in frequency) oscillates no faster (in time) than its fastest Fourier component. However, a band-limited super-oscillatory function may oscillate arbitrarily fast in regions of relatively low intensity. In optics, this means that we can create an arbitrarily small hotspot at the focus of a lens using engineered interference of light. However, super-oscillatory hotspots are necessarily surrounded by sidebands that contain some fraction of the optical power – trading efficiency for resolution. We replace the objective in a confocal microscope with a super-oscillatory lens and use the confocal pinhole to reject the light scattered from the sidebands. The resolution of the image is determined by the size of the super-oscillatory hotspot.

We have developed a super-oscillatory system to image unlabelled cells at super-resolution and high speed. To do this we combine our super-oscillatory microscope with advanced polarisation-contrast imaging. The instrument is a modification of a standard confocal microscope, with two key components: spatial light modulators to shape the laser beam entering the microscope, and a liquid crystal panel to control the input polarisation. We capture four differently-polarised super-resolved images of the sample and then calculate the anisotropy and orientation angle of each pixel. This highlights those parts of a cell with significant molecular structuring, such as actin filaments, microtubules, and even protein-enriched lipid bilayers such as vesicles and cell membranes.

We have applied this to a number of systems showing it is able to reveal new levels of information in living and moving biological samples.
0006-3495
186a
Rogers, Edward
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Quraishe, Shmma
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Chad, John
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Newman, Tracey
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Zheludev, Nikolai
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Smith, P.J.S.
003de469-9420-4f12-8f0e-8e8d76d28d6c
Rogers, Edward
b92cc8ab-0d91-4b2e-b5c7-8a2f490a36a2
Quraishe, Shmma
31673f5e-736f-4849-ba79-6e078cbd2cb2
Chad, John
d220e55e-3c13-4d1d-ae9a-1cfae8ccfbe1
Newman, Tracey
322290cb-2e9c-445d-a047-00b1bea39a25
Zheludev, Nikolai
32fb6af7-97e4-4d11-bca6-805745e40cc6
Smith, P.J.S.
003de469-9420-4f12-8f0e-8e8d76d28d6c

Rogers, Edward, Quraishe, Shmma, Chad, John, Newman, Tracey, Zheludev, Nikolai and Smith, P.J.S. (2017) New super-oscillatory technology for unlabelled super-resolution cellular imaging with polarisation contrast. Biophysical Journal, 112 (3, Supplement 1), 186a. (doi:10.1016/j.bpj.2016.11.1031).

Record type: Meeting abstract

Abstract

Super-resolution microscopy is already showing huge benefits across the biosciences, but all widely-used techniques require the addition of fluorescent probes. We have demonstrated optical-super-resolution imaging in unlabelled living cells, using the phenomenon of super-oscillation.

Super-oscillation is originally a mathematical phenomenon, first described in quantum mechanics. It is widely accepted that any function that is band-limited (in frequency) oscillates no faster (in time) than its fastest Fourier component. However, a band-limited super-oscillatory function may oscillate arbitrarily fast in regions of relatively low intensity. In optics, this means that we can create an arbitrarily small hotspot at the focus of a lens using engineered interference of light. However, super-oscillatory hotspots are necessarily surrounded by sidebands that contain some fraction of the optical power – trading efficiency for resolution. We replace the objective in a confocal microscope with a super-oscillatory lens and use the confocal pinhole to reject the light scattered from the sidebands. The resolution of the image is determined by the size of the super-oscillatory hotspot.

We have developed a super-oscillatory system to image unlabelled cells at super-resolution and high speed. To do this we combine our super-oscillatory microscope with advanced polarisation-contrast imaging. The instrument is a modification of a standard confocal microscope, with two key components: spatial light modulators to shape the laser beam entering the microscope, and a liquid crystal panel to control the input polarisation. We capture four differently-polarised super-resolved images of the sample and then calculate the anisotropy and orientation angle of each pixel. This highlights those parts of a cell with significant molecular structuring, such as actin filaments, microtubules, and even protein-enriched lipid bilayers such as vesicles and cell membranes.

We have applied this to a number of systems showing it is able to reveal new levels of information in living and moving biological samples.

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Accepted/In Press date: 1 January 2017
e-pub ahead of print date: 3 February 2017
Published date: 14 February 2017
Venue - Dates: 61st Annual Meeting of the Biophysical Society, New Orleans, United States, 2017-02-11 - 2017-02-15
Organisations: Optoelectronics Research Centre, Institute for Life Sciences, Biomedicine, Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 408124
URI: https://eprints.soton.ac.uk/id/eprint/408124
ISSN: 0006-3495
PURE UUID: 815c204c-4db4-43b1-b347-90631c8c5182
ORCID for John Chad: ORCID iD orcid.org/0000-0001-6442-4281
ORCID for Tracey Newman: ORCID iD orcid.org/0000-0002-3727-9258
ORCID for Nikolai Zheludev: ORCID iD orcid.org/0000-0002-1013-6636
ORCID for P.J.S. Smith: ORCID iD orcid.org/0000-0003-4400-6853

Catalogue record

Date deposited: 12 May 2017 04:03
Last modified: 10 Dec 2019 01:59

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Contributors

Author: Edward Rogers
Author: Shmma Quraishe
Author: John Chad ORCID iD
Author: Tracey Newman ORCID iD
Author: P.J.S. Smith ORCID iD

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