A Microfluidic Platform for Tracking Individual Cell Dynamics during an Unperturbed Nutrients Exhaustion.
Fiche publication
Date publication
juillet 2022
Journal
Bio-protocol
Auteurs
Membres identifiés du Cancéropôle Est :
Dr CHARVIN Gilles
Tous les auteurs :
Aspert T, Jacquel B, Charvin G
Lien Pubmed
Résumé
Microorganisms have evolved adaptive strategies to respond to the autonomous degradation of their environment. Indeed, a growing culture progressively exhausts nutrients from its media and modifies its composition. Yet, how single cells react to these modifications remains difficult to study since it requires population-scale growth experiments to allow cell proliferation to have a collective impact on the environment, while monitoring the same individuals exposed to this environment for days. For this purpose, we have previously described an integrated microfluidic pipeline, based on continuous separation of the cells from the media and subsequent perfusion of the filtered media in an observation chamber containing isolated single cells. Here, we provide a detailed protocol to implement this methodology, including the setting up of the microfluidic system and the processing of timelapse images.
Mots clés
Cell fate heterogeneities , Cytosolic pH , Diauxic shift , Metabolic state , Microfluidic-based time-lapse microscopy , Quiescence entry , Single-cell ecology
Référence
Bio Protoc. 2022 07 20;12(14):