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Efficient Illumination for Microsecond Tracking Microscopy

The possibility to observe microsecond dynamics at the sub-micron scale, opened by recent technological advances in fast camera sensors, will affect many biophysical studies based on particle tracking in optical microscopy. A main limiting factor for further development of fast video microscopy rema...

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Detalles Bibliográficos
Autores principales: Dulin, David, Barland, Stephane, Hachair, Xavier, Pedaci, Francesco
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4175081/
https://www.ncbi.nlm.nih.gov/pubmed/25251462
http://dx.doi.org/10.1371/journal.pone.0107335
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author Dulin, David
Barland, Stephane
Hachair, Xavier
Pedaci, Francesco
author_facet Dulin, David
Barland, Stephane
Hachair, Xavier
Pedaci, Francesco
author_sort Dulin, David
collection PubMed
description The possibility to observe microsecond dynamics at the sub-micron scale, opened by recent technological advances in fast camera sensors, will affect many biophysical studies based on particle tracking in optical microscopy. A main limiting factor for further development of fast video microscopy remains the illumination of the sample, which must deliver sufficient light to the camera to allow microsecond exposure times. Here we systematically compare the main illumination systems employed in holographic tracking microscopy, and we show that a superluminescent diode and a modulated laser diode perform the best in terms of image quality and acquisition speed, respectively. In particular, we show that the simple and inexpensive laser illumination enables less than [Image: see text]s camera exposure time at high magnification on a large field of view without coherence image artifacts, together with a good hologram quality that allows nm-tracking of microscopic beads to be performed. This comparison of sources can guide in choosing the most efficient illumination system with respect to the specific application.
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spelling pubmed-41750812014-10-02 Efficient Illumination for Microsecond Tracking Microscopy Dulin, David Barland, Stephane Hachair, Xavier Pedaci, Francesco PLoS One Research Article The possibility to observe microsecond dynamics at the sub-micron scale, opened by recent technological advances in fast camera sensors, will affect many biophysical studies based on particle tracking in optical microscopy. A main limiting factor for further development of fast video microscopy remains the illumination of the sample, which must deliver sufficient light to the camera to allow microsecond exposure times. Here we systematically compare the main illumination systems employed in holographic tracking microscopy, and we show that a superluminescent diode and a modulated laser diode perform the best in terms of image quality and acquisition speed, respectively. In particular, we show that the simple and inexpensive laser illumination enables less than [Image: see text]s camera exposure time at high magnification on a large field of view without coherence image artifacts, together with a good hologram quality that allows nm-tracking of microscopic beads to be performed. This comparison of sources can guide in choosing the most efficient illumination system with respect to the specific application. Public Library of Science 2014-09-24 /pmc/articles/PMC4175081/ /pubmed/25251462 http://dx.doi.org/10.1371/journal.pone.0107335 Text en © 2014 Dulin et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Dulin, David
Barland, Stephane
Hachair, Xavier
Pedaci, Francesco
Efficient Illumination for Microsecond Tracking Microscopy
title Efficient Illumination for Microsecond Tracking Microscopy
title_full Efficient Illumination for Microsecond Tracking Microscopy
title_fullStr Efficient Illumination for Microsecond Tracking Microscopy
title_full_unstemmed Efficient Illumination for Microsecond Tracking Microscopy
title_short Efficient Illumination for Microsecond Tracking Microscopy
title_sort efficient illumination for microsecond tracking microscopy
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4175081/
https://www.ncbi.nlm.nih.gov/pubmed/25251462
http://dx.doi.org/10.1371/journal.pone.0107335
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