Preparation, imaging, and quantification of bacterial surface motility assays.

Abstract : Bacterial surface motility, such as swarming, is commonly examined in the laboratory using plate assays that necessitate specific concentrations of agar and sometimes inclusion of specific nutrients in the growth medium. The preparation of such explicit media and surface growth conditions serves to provide the favorable conditions that allow not just bacterial growth but coordinated motility of bacteria over these surfaces within thin liquid films. Reproducibility of swarm plate and other surface motility plate assays can be a major challenge. Especially for more "temperate swarmers" that exhibit motility only within agar ranges of 0.4%-0.8% (wt/vol), minor changes in protocol or laboratory environment can greatly influence swarm assay results. "Wettability", or water content at the liquid-solid-air interface of these plate assays, is often a key variable to be controlled. An additional challenge in assessing swarming is how to quantify observed differences between any two (or more) experiments. Here we detail a versatile two-phase protocol to prepare and image swarm assays. We include guidelines to circumvent the challenges commonly associated with swarm assay media preparation and quantification of data from these assays. We specifically demonstrate our method using bacteria that express fluorescent or bioluminescent genetic reporters like green fluorescent protein (GFP), luciferase (lux operon), or cellular stains to enable time-lapse optical imaging. We further demonstrate the ability of our method to track competing swarming species in the same experiment.
Type de document :
Article dans une revue
Journal of visualized experiments : JoVE, JoVE, 2015, 〈10.3791/52338〉
Liste complète des métadonnées

Littérature citée [15 références]  Voir  Masquer  Télécharger
Contributeur : Michel Courcelles <>
Soumis le : lundi 8 août 2016 - 15:33:40
Dernière modification le : jeudi 7 février 2019 - 16:56:49


Publication financée par une institution


Distributed under a Creative Commons Paternité - Pas d'utilisation commerciale - Pas de modification 4.0 International License




Nydia Morales-Soto, Morgen E Anyan, Anne E Mattingly, Chinedu S Madukoma, Cameron W Harvey, et al.. Preparation, imaging, and quantification of bacterial surface motility assays.. Journal of visualized experiments : JoVE, JoVE, 2015, 〈10.3791/52338〉. 〈pasteur-01352584〉



Consultations de la notice


Téléchargements de fichiers