Super-resolution imaging of bacteria in a microfluidics device.

Bacteria have evolved complex, highly-coordinated, multi-component cellular engines to achieve high degrees of efficiency, accuracy, adaptability, and redundancy. Super-resolution fluorescence microscopy methods are ideally suited to investigate the internal composition, architecture, and dynamics o...

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Autores principales:	Diego I Cattoni, Jean-Bernard Fiche, Alessandro Valeri, Tâm Mignot, Marcelo Nöllmann
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9e9b511157064319856e1e424c4efd4e
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spelling	oai:doaj.org-article:9e9b511157064319856e1e424c4efd4e2021-11-18T08:50:50ZSuper-resolution imaging of bacteria in a microfluidics device.1932-620310.1371/journal.pone.0076268https://doaj.org/article/9e9b511157064319856e1e424c4efd4e2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24146850/?tool=EBIhttps://doaj.org/toc/1932-6203Bacteria have evolved complex, highly-coordinated, multi-component cellular engines to achieve high degrees of efficiency, accuracy, adaptability, and redundancy. Super-resolution fluorescence microscopy methods are ideally suited to investigate the internal composition, architecture, and dynamics of molecular machines and large cellular complexes. These techniques require the long-term stability of samples, high signal-to-noise-ratios, low chromatic aberrations and surface flatness, conditions difficult to meet with traditional immobilization methods. We present a method in which cells are functionalized to a microfluidics device and fluorophores are injected and imaged sequentially. This method has several advantages, as it permits the long-term immobilization of cells and proper correction of drift, avoids chromatic aberrations caused by the use of different filter sets, and allows for the flat immobilization of cells on the surface. In addition, we show that different surface chemistries can be used to image bacteria at different time-scales, and we introduce an automated cell detection and image analysis procedure that can be used to obtain cell-to-cell, single-molecule localization and dynamic heterogeneity as well as average properties at the super-resolution level.Diego I CattoniJean-Bernard FicheAlessandro ValeriTâm MignotMarcelo NöllmannPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 10, p e76268 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Diego I Cattoni Jean-Bernard Fiche Alessandro Valeri Tâm Mignot Marcelo Nöllmann Super-resolution imaging of bacteria in a microfluidics device.
description	Bacteria have evolved complex, highly-coordinated, multi-component cellular engines to achieve high degrees of efficiency, accuracy, adaptability, and redundancy. Super-resolution fluorescence microscopy methods are ideally suited to investigate the internal composition, architecture, and dynamics of molecular machines and large cellular complexes. These techniques require the long-term stability of samples, high signal-to-noise-ratios, low chromatic aberrations and surface flatness, conditions difficult to meet with traditional immobilization methods. We present a method in which cells are functionalized to a microfluidics device and fluorophores are injected and imaged sequentially. This method has several advantages, as it permits the long-term immobilization of cells and proper correction of drift, avoids chromatic aberrations caused by the use of different filter sets, and allows for the flat immobilization of cells on the surface. In addition, we show that different surface chemistries can be used to image bacteria at different time-scales, and we introduce an automated cell detection and image analysis procedure that can be used to obtain cell-to-cell, single-molecule localization and dynamic heterogeneity as well as average properties at the super-resolution level.
format	article
author	Diego I Cattoni Jean-Bernard Fiche Alessandro Valeri Tâm Mignot Marcelo Nöllmann
author_facet	Diego I Cattoni Jean-Bernard Fiche Alessandro Valeri Tâm Mignot Marcelo Nöllmann
author_sort	Diego I Cattoni
title	Super-resolution imaging of bacteria in a microfluidics device.
title_short	Super-resolution imaging of bacteria in a microfluidics device.
title_full	Super-resolution imaging of bacteria in a microfluidics device.
title_fullStr	Super-resolution imaging of bacteria in a microfluidics device.
title_full_unstemmed	Super-resolution imaging of bacteria in a microfluidics device.
title_sort	super-resolution imaging of bacteria in a microfluidics device.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/9e9b511157064319856e1e424c4efd4e
work_keys_str_mv	AT diegoicattoni superresolutionimagingofbacteriainamicrofluidicsdevice AT jeanbernardfiche superresolutionimagingofbacteriainamicrofluidicsdevice AT alessandrovaleri superresolutionimagingofbacteriainamicrofluidicsdevice AT tammignot superresolutionimagingofbacteriainamicrofluidicsdevice AT marcelonollmann superresolutionimagingofbacteriainamicrofluidicsdevice
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Super-resolution imaging of bacteria in a microfluidics device.

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