Live Cell Imaging of Bacillus subtilis and Streptococcus pneumoniae using Automated Time-lapse Microscopy.

Détails

ID Serval
serval:BIB_1B1D3D338AC3
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Live Cell Imaging of Bacillus subtilis and Streptococcus pneumoniae using Automated Time-lapse Microscopy.
Périodique
Journal of Visualized Experiments
Auteur(s)
de Jong I.G., Beilharz K., Kuipers O.P., Veening J.W.
ISSN
1940-087X (Electronic)
ISSN-L
1940-087X
Statut éditorial
Publié
Date de publication
2011
Numéro
53
Pages
UNSP e3145
Langue
anglais
Résumé
During the last few years scientists became increasingly aware that average data obtained from microbial population based experiments are not representative of the behavior, status or phenotype of single cells. Due to this new insight the number of single cell studies rises continuously (for recent reviews see (1,2,3)). However, many of the single cell techniques applied do not allow monitoring the development and behavior of one specific single cell in time (e.g. flow cytometry or standard microscopy). Here, we provide a detailed description of a microscopy method used in several recent studies (4, 5, 6, 7), which allows following and recording (fluorescence of) individual bacterial cells of Bacillus subtilis and Streptococcus pneumoniae through growth and division for many generations. The resulting movies can be used to construct phylogenetic lineage trees by tracing back the history of a single cell within a population that originated from one common ancestor. This time-lapse fluorescence microscopy method cannot only be used to investigate growth, division and differentiation of individual cells, but also to analyze the effect of cell history and ancestry on specific cellular behavior. Furthermore, time-lapse microscopy is ideally suited to examine gene expression dynamics and protein localization during the bacterial cell cycle. The method explains how to prepare the bacterial cells and construct the microscope slide to enable the outgrowth of single cells into a microcolony. In short, single cells are spotted on a semi-solid surface consisting of growth medium supplemented with agarose on which they grow and divide under a fluorescence microscope within a temperature controlled environmental chamber. Images are captured at specific intervals and are later analyzed using the open source software ImageJ.
Mots-clé
Bacillus subtilis/cytology, Bacteriological Techniques/methods, Microscopy, Fluorescence/methods, Streptococcus pneumoniae/cytology
Pubmed
Web of science
Open Access
Oui
Création de la notice
11/10/2016 15:30
Dernière modification de la notice
20/08/2019 12:51
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