title: Space- and time-resolved protein dynamics in single bacterial cells observed on a chip.
abstract: Life cell imaging of bacterial cells over long times is very challenging because of the small dimensions and the need for a liquid environment assuring cell viability. In order to obtain space- and time-resolved information about protein dynamics, high resolution time-lapse fluorescence images (TLFI) of single bacterial cells were recorded in a poly(dimethylsiloxane) (PDMS) microfluidic chip. A new gradient coating technique was applied to ensure cell loading. As a proof-of-concept, we monitored the evenly distributed cytoplasmic protein GcrA as well as the asymmetric localization of the DivK protein in cells of S. meliloti over at least two division cycles. Localization of DivK was characterized by dividing each bacterial cell into 4 sections with dimensions closely above the optical limit of resolution. This approach of generating spatio-temporal resolved information of protein dynamics in single bacterial cells is applicable to many problems.
authors.affiliation: Experimental Biophysics & Applied Nanoscience, Bielefeld University, Universitaetsstr. 25, 33615 Bielefeld, Germany.
authors.forename: Dominik
authors.initials: D
authors.lastname: Greif
authors.forename: Nataliya
authors.initials: N
authors.lastname: Pobigaylo
authors.forename: Benjamin
authors.initials: B
authors.lastname: Frage
authors.forename: Anke
authors.initials: A
authors.lastname: Becker
authors.forename: Jan
authors.initials: J
authors.lastname: Regtmeier
authors.forename: Dario
authors.initials: D
authors.lastname: Anselmetti
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D002453:Cell Cycle / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D044085:Microfluidics / Q000379:methods*; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence / Q000379:methods; D011506:Proteins / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D016962:Sinorhizobium meliloti / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D011506:Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins