title: Imaging flow cytometry analysis of intracellular pathogens.
abstract: Imaging flow cytometry has been applied to address questions in infection biology, in particular, infections induced by intracellular pathogens. This methodology, which utilizes specialized analytic software makes it possible to analyze hundreds of quantified features for hundreds of thousands of individual cellular or subcellular events in a single experiment. Imaging flow cytometry analysis of host cell-pathogen interaction can thus quantitatively addresses a variety of biological questions related to intracellular infection, including cell counting, internalization score, and subcellular patterns of co-localization. Here, we provide an overview of recent achievements in the use of fluorescently labeled prokaryotic or eukaryotic pathogens in human cellular infections in analysis of host-pathogen interactions. Specifically, we give examples of Imagestream-based analysis of cell lines infected with Toxoplasma gondii or Mycobacterium tuberculosis. Furthermore, we illustrate the capabilities of imaging flow cytometry using a combination of standard IDEAS™ software and the more recently developed Feature Finder algorithm, which is capable of identifying statistically significant differences between researcher-defined image galleries. We argue that the combination of imaging flow cytometry with these software platforms provides a powerful new approach to understanding host control of intracellular pathogens.
authors.affiliation: Program in Cellular and Molecular Medicine, Boston Children's Hospital, Harvard Medical School, United States; Department of Pediatrics, Harvard Medical School, United States.
authors.forename: Viraga
authors.initials: V
authors.lastname: Haridas
authors.affiliation: Program in Cellular and Molecular Medicine, Boston Children's Hospital, Harvard Medical School, United States; Department of Pediatrics, Harvard Medical School, United States.
authors.forename: Shahin
authors.initials: S
authors.lastname: Ranjbar
authors.affiliation: School of Science and Technology, Nazarbayev University, Kazakhstan; A.N. Belozersky Institute of Physico-Chemical Biology, M.V. Lomonosov Moscow State University, Russia; Department of Cell Biology and Histology, M.V. Lomonosov Moscow State University, Russia.
authors.forename: Ivan A
authors.initials: IA
authors.lastname: Vorobjev
authors.affiliation: Program in Cellular and Molecular Medicine, Boston Children's Hospital, Harvard Medical School, United States; Department of Pediatrics, Harvard Medical School, United States. Electronic address: Anne.Goldfeld@childrens.harvard.edu.
authors.forename: Anne E
authors.initials: AE
authors.lastname: Goldfeld
authors.affiliation: Program in Cellular and Molecular Medicine, Boston Children's Hospital, Harvard Medical School, United States; Department of Pediatrics, Harvard Medical School, United States; School of Science and Technology, Nazarbayev University, Kazakhstan. Electronic address: natasha.barteneva@childrens.harvard.edu.
authors.forename: Natasha S
authors.initials: NS
authors.lastname: Barteneva
mesh_terms: D000465:Algorithms; D000943:Antigens, Differentiation / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D054884:Host-Pathogen Interactions; D006801:Humans; D019044:Image Cytometry / Q000295:instrumentation / Q000379:methods*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000378:metabolism* / Q000648:ultrastructure; D010587:Phagocytosis; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D012984:Software; D013194:Staining and Labeling / Q000379:methods; D000074084:THP-1 Cells; D014122:Toxoplasma / Q000378:metabolism* / Q000648:ultrastructure; D000097573:Red Fluorescent Protein
chemical_list: D000943:Antigens, Differentiation; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D034741:RNA, Small Interfering; D000097573:Red Fluorescent Protein; C062010:leu-13 antigen
keywords: Cellular heterogeneity; Colocalization; Feature Finder; Fluorescent protein; Imaging flow cytometry; Intracellular pathogen; Mycobacteria tuberculosis; Phagosome maturation; Rab5; Rab7; Toxoplasma gondii