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Nano Lett. 2015 May 13;15(5):2928-32. doi: 10.1021/nl504710z. Epub 2015 Apr 17.

Live-Cell STED Microscopy with Genetically Encoded Biosensor.

Author information

1
†Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.
2
‡Nizhny Novgorod State Medical Academy, Nizhny Novgorod 603005, Russia.
3
§EMBL Heidelberg, Heidelberg 69117, Germany.

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.

KEYWORDS:

HyPer; STED; biosensor; hydrogen peroxide; microscopy; super-resolution imaging

PMID:
25871892
DOI:
10.1021/nl504710z
[Indexed for MEDLINE]

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