Fast live simultaneous multiwavelength four-dimensional optical microscopy

Peter M. Carlton; Jérôme Boulanger; Charles Kervrann; Jean-Baptiste Sibarita; Jean Salamero; Et Al., John W. Sedat

inria-00540978, version 1

Fast live simultaneous multiwavelength four-dimensional optical microscopy

Peter M. Carlton^c^, 1, 2, Jérôme Boulanger^a^, 3, Charles Kervrann () ^4, 5, Jean-Baptiste Sibarita^b^, 3, 6, 7, Jean Salamero ()^b^, 3, 7, Et Al., John W. Sedat^c^, 1, 2, 8

Proc Natl Acad Sci USA 107, 37 (2010) 16016-16022

Résumé : Live fluorescence microscopy has the unique capability to probe dynamic processes, linking molecular components and their localization with function. A key goal of microscopy is to increase spatial and temporal resolution while simultaneously permitting identification of multiple specific components. We demonstrate a new microscope platform, OMX, that enables subsecond, multicolor four-dimensional data acquisition and also provides access to subdiffraction structured illumination imaging. Using this platform to image chromosome movement during a complete yeast cell cycle at one 3D image stack per second reveals an unexpected degree of photosensitivity of fluorophore-containing cells. To avoid perturbation of cell division, excitation levels had to be attenuated between 100 and 10,000× below the level normally used for imaging. We show that an image denoising algorithm that exploits redundancy in the image sequence over space and time allows recovery of biological information from the low light level noisy images while maintaining full cell viability with no fading.

a – Institut Curie, Centre de Recherche, 12 rue Lhomond, 75005, Paris
b – CNRS
c – UCSF
1 : The Keck Center for Advanced Microscopy
University of California
2 : Department of Biochemistry and Biophysics [San Francisco]
University of California
3 : Cell and Tissue Imaging Facilities, UMR 144 CNRS (Pict-IBiSA)
Institut Curie
4 : INRA - Mathématiques et Informatique Appliquées (Unité MIAJ)
Institut national de la recherche agronomique (INRA)
5 : SERPICO (INRIA)
INRIA
6 : Physiologie cellulaire de la synapse (PCS)
CNRS : UMR5091 – Institut François Magendie – Université Victor Segalen - Bordeaux II
7 : Molecular mechanisms of intracellular transport, UMR 144 CNRS
Institut Curie
8 : Howard Hughes Medical Institute (HHMI)
Howard Hugues Medical Institute

inria-00540978, version 1
http://hal.inria.fr/inria-00540978
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Contributeur : Charles Kervrann <>
Soumis le : Lundi 29 Novembre 2010, 15:39:24
Dernière modification le : Jeudi 31 Janvier 2013, 15:00:38

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DOI : 10.1073/pnas.1004037107

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%% inria-00540978, version 1
%% http://hal.inria.fr/inria-00540978
@article{carlton:inria-00540978,
    hal_id = {inria-00540978},
    url = {http://hal.inria.fr/inria-00540978},
    title = {{Fast live simultaneous multiwavelength four-dimensional optical microscopy}},
    author = {Carlton, Peter M. and Boulanger, J{\'e}r{\^o}me and Kervrann, Charles and Sibarita, Jean-Baptiste and Salamero, Jean and Sedat, Et Al., John W.},
    abstract = {{Live fluorescence microscopy has the unique capability to probe dynamic processes, linking molecular components and their localization with function. A key goal of microscopy is to increase spatial and temporal resolution while simultaneously permitting identification of multiple specific components. We demonstrate a new microscope platform, OMX, that enables subsecond, multicolor four-dimensional data acquisition and also provides access to subdiffraction structured illumination imaging. Using this platform to image chromosome movement during a complete yeast cell cycle at one 3D image stack per second reveals an unexpected degree of photosensitivity of fluorophore-containing cells. To avoid perturbation of cell division, excitation levels had to be attenuated between 100 and 10,000× below the level normally used for imaging. We show that an image denoising algorithm that exploits redundancy in the image sequence over space and time allows recovery of biological information from the low light level noisy images while maintaining full cell viability with no fading.}},
    language = {Anglais},
    affiliation = {The Keck Center for Advanced Microscopy , Department of Biochemistry and Biophysics [San Francisco] , Cell and Tissue Imaging Facilities, UMR 144 CNRS (Pict-IBiSA) , INRA - Math{\'e}matiques et Informatique Appliqu{\'e}es - Unit{\'e} MIAJ , SERPICO - INRIA , Physiologie cellulaire de la synapse - PCS , Molecular mechanisms of intracellular transport, UMR 144 CNRS , Howard Hughes Medical Institute - HHMI},
    publisher = {PNAS},
    pages = {16016-16022},
    journal = {Proc Natl Acad Sci USA},
    volume = {107},
    number = {37 },
    audience = {internationale },
    doi = {10.1073/pnas.1004037107 },
    year = {2010},
    month = Sep,
}

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