title: A confocal microscopy image analysis method to measure adhesion and internalization of Pseudomonas aeruginosa multicellular structures into epithelial cells.
abstract: Formation of multicellular structures such as biofilms is an important feature in the physiopathology of many disease-causing bacteria. We recently reported that Pseudomonas aeruginosa adheres to epithelial cells rapidly forming early biofilm-like aggregates, which can then be internalized into cells. Conventional methods to measure adhesion/internalization, such as dilution plating for total cell-associated or antibiotic protected bacteria, do not distinguish between single and aggregated bacteria. We report a procedure that combining double bacteria labeling, confocal microscopy and image analysis allows identification and quantification of the number of adhered and internalized bacteria distinguishing between single and aggregated bacterial cells. A plugin for Fiji to automatically perform these procedures has been generated.
authors.forename: Paola
authors.initials: P
authors.lastname: Lepanto
authors.forename: Federico
authors.initials: F
authors.lastname: Lecumberry
authors.forename: Jéssica
authors.initials: J
authors.lastname: Rossello
authors.forename: Arlinet
authors.initials: A
authors.lastname: Kierbel
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion; D018441:Biofilms; D004285:Dogs; D004847:Epithelial Cells / Q000382:microbiology*; D054884:Host-Pathogen Interactions; D007091:Image Processing, Computer-Assisted; D061985:Madin Darby Canine Kidney Cells; D018613:Microscopy, Confocal / Q000379:methods*; D011550:Pseudomonas aeruginosa / Q000472:pathogenicity*; D012984:Software