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Standing-wave-excited multiplanar fluorescence in a laser scanning microscope reveals 3D information on red blood cells

Standing-wave-excited multiplanar fluorescence in a laser scanning microscope reveals 3D information on red blood cells
Standing-wave-excited multiplanar fluorescence in a laser scanning microscope reveals 3D information on red blood cells
Standing-wave excitation of fluorescence is highly desirable in optical microscopy because it improves the axial resolution. We demonstrate here that multiplanar excitation of fluorescence by a standing wave can be produced in a single-spot laser scanning microscope by placing a plane reflector close to the specimen. We report here a variation in the intensity of fluorescence of successive planes related to the Stokes shift of the dye. We show by the use of dyes specific for the cell membrane how standing-wave excitation can be exploited to generate precise contour maps of the surface membrane of red blood cells, with an axial resolution of ?90?nm. The method, which requires only the addition of a plane mirror to an existing confocal laser scanning microscope, may well prove useful in studying diseases which involve the red cell membrane, such as malaria.
fluorescence imaging, biological fluorescence, optical physics
1-7
Amor, Rumelo
bd8b5ddf-7d00-44fc-880d-507e3293a588
Mahajan, Sumeet
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Amos, William Bradshaw
a2b96c03-3fec-48a3-8ee0-d41227897e77
McConnell, Gail
09172b64-80d3-47fd-ac78-de7b018873e0
Amor, Rumelo
bd8b5ddf-7d00-44fc-880d-507e3293a588
Mahajan, Sumeet
b131f40a-479e-4432-b662-19d60d4069e9
Amos, William Bradshaw
a2b96c03-3fec-48a3-8ee0-d41227897e77
McConnell, Gail
09172b64-80d3-47fd-ac78-de7b018873e0

Amor, Rumelo, Mahajan, Sumeet, Amos, William Bradshaw and McConnell, Gail (2014) Standing-wave-excited multiplanar fluorescence in a laser scanning microscope reveals 3D information on red blood cells. Scientific Reports, 4 (7359), 1-7. (doi:10.1038/srep07359).

Record type: Article

Abstract

Standing-wave excitation of fluorescence is highly desirable in optical microscopy because it improves the axial resolution. We demonstrate here that multiplanar excitation of fluorescence by a standing wave can be produced in a single-spot laser scanning microscope by placing a plane reflector close to the specimen. We report here a variation in the intensity of fluorescence of successive planes related to the Stokes shift of the dye. We show by the use of dyes specific for the cell membrane how standing-wave excitation can be exploited to generate precise contour maps of the surface membrane of red blood cells, with an axial resolution of ?90?nm. The method, which requires only the addition of a plane mirror to an existing confocal laser scanning microscope, may well prove useful in studying diseases which involve the red cell membrane, such as malaria.

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srep_rev_v3.pdf - Accepted Manuscript
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More information

Accepted/In Press date: 19 November 2014
Published date: 8 December 2014
Keywords: fluorescence imaging, biological fluorescence, optical physics
Organisations: Institute for Life Sciences, Organic Chemistry: SCF

Identifiers

Local EPrints ID: 372885
URI: http://eprints.soton.ac.uk/id/eprint/372885
PURE UUID: a632a305-13e6-4e87-a219-72de7eddab5e
ORCID for Sumeet Mahajan: ORCID iD orcid.org/0000-0001-8923-6666

Catalogue record

Date deposited: 24 Dec 2014 14:40
Last modified: 21 Nov 2021 02:59

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Contributors

Author: Rumelo Amor
Author: Sumeet Mahajan ORCID iD
Author: William Bradshaw Amos
Author: Gail McConnell

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