title: Visualization of intracellular hydrogen peroxide with HyPer, a genetically encoded fluorescent probe.
abstract: The fluorescent sensor HyPer allows monitoring of intracellular H2O2 levels with a high degree of sensitivity and specificity. Here, we provide a detailed protocol of ratiometric imaging of H2O2 produced by cells during phagocytosis, including instructions for experiments on different commercial confocal systems, namely, Leica SP2, Leica SP5, and Carl Zeiss LSM, as well as wide-field Leica 6000 microscope. The general experimental scheme is easily adaptable for imaging H2O2 production by various cell types under a variety of conditions.
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, RAS, Moscow, Russia.
authors.forename: Nataliya M
authors.initials: NM
authors.lastname: Mishina
authors.forename: Kseniya N
authors.initials: KN
authors.lastname: Markvicheva
authors.forename: Dmitry S
authors.initials: DS
authors.lastname: Bilan
authors.forename: Mikhail E
authors.initials: ME
authors.lastname: Matlashov
authors.forename: Marina V
authors.initials: MV
authors.lastname: Shirmanova
authors.forename: David
authors.initials: D
authors.lastname: Liebl
authors.forename: Carsten
authors.initials: C
authors.lastname: Schultz
authors.forename: Sergey
authors.initials: S
authors.lastname: Lukyanov
authors.forename: Vsevolod V
authors.initials: VV
authors.lastname: Belousov
mesh_terms: D000818:Animals; D002460:Cell Line; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis* / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods*; D061848:Optical Imaging / Q000379:methods; D010587:Phagocytosis; D014162:Transfection / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D006861:Hydrogen Peroxide