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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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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. |
format | Online Article Text |
id | pubmed-4175081 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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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