title: Testing chemical and genetic Modulators in Mycobacterium tuberculosis infected cells using phenotypic assays.
abstract: Mycobacterium tuberculosis is able to colonize host cells, and it is now well admitted that the intracellular stage of the bacteria contributes to tuberculosis pathogenesis as well as to making it a persistent infection. There is still limited understanding on how the tubercle bacillus colonizes the cell and what are the factors impacting on its intracellular persistence. Recent advances in imaging technique allow rapid quantification of biological objects in complex environments. Furthermore, M. tuberculosis is a microorganism that is particularly genetically tractable and that tolerates the expression of heterologous fluorescent proteins. Thus, the intracellular distribution of M. tuberculosis expressing fluorescent proteins can be easily quantified by the use of confocal microscopy. Here we describe high-content/high-throughput imaging methods that enable tracking the bacillus inside host settings, taking into account the heterogeneity of colonization.
authors.affiliation: Institut Pasteur de Lille, University of Lille Inserm U1019, CNRS UMR8204, Institut Pasteur de Lille, 1, rue du Professeur Calmette, BP245, Lille Cedex, 59019, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Delorme
authors.forename: Ok-Ryul
authors.initials: OR
authors.lastname: Song
authors.forename: Alain
authors.initials: A
authors.lastname: Baulard
authors.forename: Priscille
authors.initials: P
authors.lastname: Brodin
mesh_terms: D000818:Animals; D000995:Antitubercular Agents / Q000378:metabolism / Q000494:pharmacology*; D018929:Cell Culture Techniques; D003713:Dendritic Cells / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism; D057166:High-Throughput Screening Assays; D006801:Humans; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics*; D010539:Permeability; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*
chemical_list: D000995:Antitubercular Agents; D034741:RNA, Small Interfering