title: Live-Cell STED Microscopy with Genetically Encoded Biosensor.
abstract: Of the various super-resolution techniques, stimulated emission depletion (STED) microscopy achieves the best temporal resolution at high spatial resolution, enabling live-cell imaging beyond the diffraction limit. However, STED and most other super-resolution imaging methods utilize a particular type of information extractable from the raw data, namely the positions of fluorophores. To expand on the use of super-resolution techniques, we report here the live-cell STED microscopy of a dynamic biosensor. Using the fluorescent H2O2 sensor HyPer2 for subdiffraction imaging, we were able not only to image filaments with superior resolution by localizing emission but also to trace H2O2 produced within living cell by monitoring brightness of the probe. STED microscopy of HyPer2 demonstrates potential utility of FP-based biosensors for super-resolution experiments in situ and in vivo.
authors: †Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.|‡Nizhny Novgorod State Medical Academy, Nizhny Novgorod 603005, Russia.
authors: Natalia M
authors: NM
authors: Mishina
authors: †Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.|‡Nizhny Novgorod State Medical Academy, Nizhny Novgorod 603005, Russia.
authors: Alexander S
authors: AS
authors: Mishin
authors: §EMBL Heidelberg, Heidelberg 69117, Germany.
authors: Yury
authors: Y
authors: Belyaev
authors: †Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.
authors: Ekaterina A
authors: EA
authors: Bogdanova
authors: †Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.|‡Nizhny Novgorod State Medical Academy, Nizhny Novgorod 603005, Russia.
authors: Sergey
authors: S
authors: Lukyanov
authors: §EMBL Heidelberg, Heidelberg 69117, Germany.
authors: Carsten
authors: C
authors: Schultz
authors: †Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.|‡Nizhny Novgorod State Medical Academy, Nizhny Novgorod 603005, Russia.
authors: Vsevolod V
authors: VV
authors: Belousov
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D003599:Cytoskeleton / Q000737:chemistry / Q000648:ultrastructure*; D005456:Fluorescent Dyes / Q000737:chemistry; D006861:Hydrogen Peroxide / Q000737:chemistry; D008856:Microscopy, Fluorescence
chemical_list: D005456:Fluorescent Dyes; D006861:Hydrogen Peroxide
keywords: HyPer; STED; biosensor; hydrogen peroxide; microscopy; super-resolution imaging