Application of Fluorescent Nanoparticles to Study Remodeling of the Endolysosomal System by Intracellular Bacteria

Autor(en): Zhang, Yuying
Krieger, Viktoria
Hensel, Michael 
Stichwörter: DOTS; endo-lysosomal system; fluorescent nanoparticles; GOLD NANOPARTICLES; HOST-CELL; Immunology; intracellular bacteria; Issue 95; labeling; MAMMALIAN-CELLS; Multidisciplinary Sciences; PATHOGENS; PROTEINS; quantitative image analysis; SALMONELLA; Science & Technology - Other Topics; tubular compartments
Erscheinungsdatum: 2015
Herausgeber: JOURNAL OF VISUALIZED EXPERIMENTS
Journal: JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Ausgabe: 95
Zusammenfassung: 
Fluorescent nanoparticles (NPs) with desirable chemical, optical and mechanical properties are promising tools to label intracellular organelles. Here, we introduce a method using gold-BSA-rhodamine NPs to label the endo-lysosomal system of eukaryotic cells and monitor manipulations of host cellular pathways by the intracellular pathogen Salmonella enterica. The NPs were readily internalized by HeLa cells and localized in late endosomes/lysosomes. Salmonella infection induced rearrangement of the vesicles and accumulation of NPs in Salmonella-induced membrane structures. We deployed the Imaris software package for quantitative analyses of confocal microscopy images. The number of objects and their size distribution in non-infected cells were distinct from the ones in Salmonella-infected cells, indicating extremely remodeling of the endolysosomal system by WT Salmonella.
ISSN: 1940087X
DOI: 10.3791/52058

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