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Deep-Red Fluorescent Organic Nanoparticles with High Brightness and Photostability for Super-Resolution in Vitro and in Vivo Imaging Using STED Nanoscopy

Yanzi Xu,Haoke Zhang,Ning Zhang,Xiaochi Wang,Dongfeng Dang,Xunan Jing,Duo Xi,Ying Hao,Ben Zhong Tang,Lingjie Meng

To achieve super-resolution imaging in biological research using stimulated emission depletion (STED) nanoscopy, organic luminescent materials and their corresponding fluorescent nanoparticles with high brightness and photostability are of great significance. Herein, donor-acceptor-typed DBTBT-4C8 bearing flexible alkyl chains was developed, not only to afford deep-red emission from 600 to 800 nm but also to obtain high fluorescent brightness with the absolute photoluminescence quantum yields of 25%. After that, well-defined and monodispersed spherical nanoparticles using DBTBT-4C8 with bright emission, excellent biocompatibility, and photostability, which can easily mix with amphipathic block polymers, were then produced for super-resolution in vitro and in vivo imaging using STED nanoscopy. The observations showed that in contrast to confocal microscopy with a full width at half-maximum (FWHM) value of ≈400 nm, superior resolution with a significantly improved FWHM value of only 100 nm was achieved in biomedical cell imaging, which was also used to reconstruct three-dimensional images of stained HeLa cells at an ultrahigh resolution. More importantly, by using the prepared fluorescent organic nanoparticles (FONPs) in STED nanoscopy, in vivo imaging in glass catfish with largely enhanced resolution was also successfully achieved, demonstrating that these developed deep-red FONPs here are highly suitable for super-resolution in vitro and in vivo imaging using STED nanoscopy.