RESOLFT nanoscopy with photoswitchable organic fluorophores

• DOI: 10.1038/srep17804
• Publication Date: 2015-12-07T10:33:33Z
• Authors: Jiwoong Kwon; Jihee Hwang; Jaewan Park; Gi Rim Han; Kyu Young Han; Seong Keun Kim
• Keywords: Male; 0303 health sciences; Light; Buffers; Carbocyanines; Article; Fluorescence; Mice; 03 medical and health sciences; Imaging, Three-Dimensional; Molecular Probes; NIH 3T3 Cells; Animals; Humans; Nanotechnology; Organic Chemicals; Dimerization; Fluorescent Dyes; Subcellular Fractions

Abstract Far-field optical nanoscopy has been widely used to image small objects with sub-diffraction-limit spatial resolution. Particularly, reversible saturable fluorescence transition (RESOLFT) photoswitchable fluorescent proteins is a powerful method for super-resolution imaging of living cells low light intensity. Here we demonstrate the first time implementation RESOLFT biological system using organic fluorophores, which are smaller in size and easier be chemically modified. With covalently-linked dye pair Cy3 Alexa647 label subcellular structures fixed by optimizing buffer parameters, our achieved resolution ~74 nm focal plane. This provides alternative intensity nanoscopy, enables probes at nanoscale