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Superb reliability and biocompatibility equip aggregation-induced emission (AIE) dots with tremendous potential for fluorescence bioimaging. However, there is still a chronic lack of design instructions of excretable and bright AIE emitters. Here, a kind of PEGylated AIE (OTPA-BBT) dots with strong absorption and extremely high second near-infrared region (NIR-II) PLQY of 13.6% is designed, and a long-aliphatic-chain design blueprint contributing to their excretion from an animal's body is proposed. Assisted by the OTPA-BBT dots with bright fluorescence beyond 1100 nm and even 1500 nm (NIR-IIb), large-depth cerebral vasculature (beyond 600 µm) as well as real-time blood flow are monitored through a thinned skull, and noninvasive NIR-IIb imaging with rich high-spatial-frequency information gives a precise presentation of gastrointestinal tract in marmosets. Importantly, after intravenous or oral administration, the definite excretion of OTPA-BBT dots from the body is demonstrated, which provides influential evidence of biosafety. © 2021 Wiley-VCH GmbH

Near-infrared (NIR) fluorescence is very important for high-contrast biological imaging of high-scattering tissues such as brain tissue. Unfortunately, commercial NIR dyes are excited usually by visible light, and their multi-photon absorption (MPA) cross-sections are small. Here, we design new co-encapsulated NIR nanoparticles (NPs) with a large three-photon (3PA) absorption cross-section. A form of aggregation-induced emission (AIE) luminogen (AIEgen), 2,3-bis(4-(diphenylamino)-[1,1-biphenyl]-4-yl) fumaronitrile (TPATCN), is introduced as the donor, and a form of NIR dye, silicon 2,3-naphthalocyanine bis-(trihexylsilyloxide) (NIR775), is adopted as the acceptor. Under the excitation of a 1550 nm fs laser, TPATCN-NIR775 NPs demonstrated a bright three-photon fluorescence centered at 785 nm. The energy transfer efficiency of the TPATCN-NIR775 NPs was calculated to be as high as 90%, which could be attributed to the good spectral overlap between the emission of TPATCN and the absorption of NIR775. By injection with TPATCN-NIR775 NPs, a vivid 3D reconstruction of mouse brain vasculature was obtained with even small blood vessels clearly visualized. The design strategy used for the co-encapsulated AIE-NIR NPs would be helpful in synthesizing more NIR probes for deep-tissue biological imaging in the future., QC 20180720

Fluorescence imaging in the spectral region beyond the conventional near-infrared biological window (700-900 nm) can theoretically afford high resolution and deep tissue penetration. Although some efforts have been devoted to developing a short-wave infrared (SWIR; 900-1700 nm) imaging modality in the past decade, long-wavelength biomedical imaging is still suboptimal owing to the unsatisfactory materials properties of SWIR fluorophores. Taking advantage of organic dots based on an aggregation-induced emission luminogen (AIEgen), herein microscopic vasculature imaging of brain and tumor is reported in living mice in the SWIR spectral region. The long-wavelength emission of AIE dots with certain brightness facilitates resolving brain capillaries with high spatial resolution (approximate to 3 mu m) and deep penetration (800 mu m). Owning to the deep penetration depth and real-time imaging capability, in vivo SWIR microscopic angiography exhibits superior resolution in monitoring blood-brain barrier damage in mouse brain, and visualizing enhanced permeability and retention effect in tumor sites. Furthermore, the AIE dots show good biocompatibility, and no noticeable abnormalities, inflammations or lesions are observed in the main organs of the mice. This work will inspire new insights on development of advanced SWIR techniques for biomedical imaging.

Currently, a serious problem obstructing the large-scale clinical applications of fluorescence technique is the shallow penetration depth. Two-photon fluorescence microscopic imaging with excitation in the longer-wavelength near-infrared (NIR) region (>1100 nm) and emission in the NIR-I region (650-950 nm) is a good choice to realize deep-tissue and high-resolution imaging. Here, we report ultradeep two-photon fluorescence bioimaging with 1300 nm NIR-II excitation and NIR-I emission (peak similar to 810 nm) based on a NIR aggregation-induced emission luminogen (AIEgen). The crab-shaped AIEgen possesses a planar core structure and several twisting phenyl/naphthyl rotators, affording both high fluorescence quantum yield and efficient two-photon activity. The organic AIE dots show high stability, good biocompatibility, and a large two-photon absorption cross section of 1.22 x 10(3) GM. Under 1300 nm NIR-II excitation, in vivo two-photon fluorescence microscopic imaging helps to reconstruct the 3D vasculature with a high spatial resolution of sub-3.5 mu m beyond the white matter (>840 mu m) and even to the hippocampus (> 960 mu m) and visualize small vessels of similar to 5 mu m as deep as 1065 mu m in mouse brain, which is among the largest penetration depths and best spatial resolution of in vivo two-photon imaging. Rational comparison with the AIE dots manifests that two-photon imaging outperforms the one-photon mode for high-resolution deep imaging. This work will inspire more sight and insight into the development of efficient NIR fluorophores for deep-tissue biomedical imaging.