De Novo Design of Aggregation-Induced Emission Luminogen for Three-Photon Fluorescence Imaging of Subcortical Structures Excited at Both NIR-III and NIR-IV Windows

Three-photon fluorescence (3PF) imaging excited at 1700 nm window is an enabling technology for visualizing deep brain structures and dynamics. Recently, the 2200 nm window has emerged as the longest excitation window suitable for deep-brain 3PF imaging. Bright fluorescent probes lay the material basis for deep-brain 3PF imaging. Among various fluorescent probes, aggregation-induced emission luminogens (AIEgens) have great potential in 3PF imaging excited at the 1700 nm window in vivo. However, to the best of knowledge, there is no AIEgens applicable to 3PF imaging excited at both the 1700 and 2200 nm windows. To readily fill this gap, here this study designs and synthesizes a novel AIEgen, namely TPE-DPTT-ICP, which generates bright 3PF signals excited at both 1700 and 2200 nm. The accordingly fabricated TPE-DPTT-ICP nanoparticles (NPs) possess excellent water dispersibility, colloidal stability, biocompatibility, photostability and large 3P action cross section, key to in vivo imaging. In mouse brain in vivo, TPE-DPTT-ICP NPs enable deep-brain 3PF imaging of subcortical structures excited at both the two windows, reaching depths of 1640 and 880 µm below the brain surface, respectively. TPE-DPTT-ICP NPs are thus a versatile material simultaneously catering to the need at two infrared optical windows with deep tissue penetration.     

Zwitterionic AIEgens: Rational Molecular Design for NIR-II Fluorescence Imaging-Guided Synergistic Phototherapy

Fluorescence imaging in the second near-infrared region (NIR-II) can penetrate tissue at centimeter depths and obtain high image fidelity. However, facile synthesis of small-molecule fluorescent photosensitizers for efficient NIR-II fluorescence imaging as well as photodynamic and photothermal combinatorial therapies is still a challenging task. Herein, a rational design and facile synthesis protocol are reported for a series of novel NIR-emissive zwitterionic luminogens with aggregation-induced emission (AIE) features for cancer phototheranostics. Consistent with the intrinsic features including long emission wavelength, effective reactive oxygen species generation, and excellent photothermal conversion efficiency (35.76%), in vitro and in vivo evaluation show that one of these presented AIE luminogens provides excellent performance in NIR-II fluorescence imaging-guided synergistic phototherapy against cancer.     

大鼠早期急性心肌缺血的双光子荧光成像及分析

双光子荧光显微成像技术具有较低的光漂白与光损伤、较佳的成像对比度、较深的穿透深度等优点。采用结扎大鼠冠状动脉左前降支的方法建立急性心肌缺血模型,通过双光子荧光显微成像技术获得不同结扎时间下大鼠左心室前壁心肌组织的双光子荧光图像。从中分析了不同结扎时间下心肌组织的形态学变化,实现对心肌组织损伤范围的确定;并对双光子荧光图像进行快速傅里叶变换(FFT),使用方向指数这一参数对心肌缺血的程度进行初步的量化评估。结果表明,双光子荧光显微成像技术结合FFT分析技术,有望在心肌缺血性疾病的诊断和治疗中实时、快速、准确地判断心肌缺血的范围和评估其损伤程度。     

Side-Chain Engineering of Aggregation-Induced Emission Molecules for Boosting Cancer Phototheranostics

The ingenious construction of versatile cancer phototheranostics involving fluorescence imaging (FLI) and photodynamic and photothermal therapies (PDT, PTT) concurrently has attracted great interest. By virtue of their inherent twisted structures and plentiful motion moieties, aggregation-induced emission luminogens (AIEgens) have been proven to be perfect templates for the development of multimodal phototheranostic systems as their diverse energy consumption pathways can be flexibly regulated through tuning the intramolecular motions. Side-chain engineering is generally accepted as a useful regulation strategy for intramolecular motions through altering the side-chain structure of the molecule, but has rarely been reported for the construction of AIE-active multimodal phototheranostics. Herein, by taking full advantage of the side-chain engineering strategy, an AIE-active multifunctional phototheranostic system (TBFT2 nanoparticles) is successfully constructed by intentionally manipulating the length of side chains. Bearing the longest alkyl chain, all of those three energy dissipation pathways including radiative decay, nonradiative thermal deactivation, and intersystem crossing process of TBFT2 are retained simultaneously and controllably in the aggregate state. In vitro and in vivo evaluations verify that TBFT2 nanoparticles perform well in terms of FLI-guided PDT and PTT synergistic cancer therapy. This study thus provides new insight into the exploration of superior versatile phototheranostics through side-chain engineering.     

A Molecular Brush Approach to Enhance Quantum Yield and Suppress Nonspecific Interactions of Conjugated Polyelectrolyte for Targeted Far‐Red/Near‐Infrared Fluorescence Cell Imaging

A red‐fluorescent conjugated polyelectrolyte (CPE, P2 ) is grafted with dense poly(ethylene glycol) (PEG) chains via click chemistry and subsequently modified with folic acid to form a molecular brush based cellular probe ( P4 ). P4 self‐assembles into a core–shell nanostructure in aqueous medium with an average size of 130 nm measured by laser light scattering. As compared to P2 , P4 possesses not only a substantially higher quantum yield (11%), but also reduced nonspecific interactions with biomolecules in aqueous medium due to the shielding effect of PEG. In conjunction with its high photostability and low cytotoxicity, utilization of P4 as a far‐red/near‐infrared cellular probe allows for effective visualization and discrimination of MCF‐7 cancer cells from NIH‐3T3 normal cells in a high contrast, selective, and nonviral manner. This study thus demonstrates a flexible molecular brush approach to overcome the intrinsic drawbacks of CPEs for advanced bioimaging applications.     

Deepening Insights into Aggregation Effect of Intermolecular Charge-Transfer Aggregates for Highly Efficient Near-Infrared Non-Doped Organic Light-Emitting Diodes over 780 nm

Though urgently needed, high-efficiency near-infrared (NIR) organic light-emitting diode (OLED) is still rare due to the energy-gap law. Formation of intermolecular charge-transfer aggregates (CTA) with nonadiabatic coupling suppression can decelerate non-radiative decay rates for high-efficiency NIR-OLEDs. However, the aggregation effect of CTA is still not fully understood, which limits the rational design of CTA. Herein, two CTA molecules with a same π-framework but different terminal substituents are developed to unveil the aggregation effect. In highly ordered crystalline states, the terminal substituents substantially affect the molecular packing motifs and intermolecular charge-transfer states, thus leading to distinct photophysical properties. In comparison, in amorphous states, these two CTA demonstrate similar photophysical behaviors and properties due to their similar molecular packing and intermolecular interactions as evidenced by molecular dynamics simulations. Importantly, the formations of amorphous CTA trigger multifunction improvements such as aggregation-induced NIR emission, aggregation-induced thermally activated delayed fluorescence, self-doping and self-host features. The non-doped OLEDs demonstrate NIR emissions centered at 788 and 803 nm, and high maximum external quantum efficiencies of 2.6% and 1.5% with small efficiency roll-off, respectively. This study provides deeper insight into the aggregation effect of CTA and lays a foundation for the development of high-efficiency NIR non-doped OLEDs.     

Robust Red Organic Nanoparticles for In Vivo Fluorescence Imaging of Cancer Cell Progression in Xenografted Zebrafish

Bright and red‐emissive organic nanoparticles (NPs) are demonstrated as promising for in vivo fluorescence imaging. However, most red organic dyes show greatly weakened or quenched emission in the aggregated state. In this work, a robust red fluorophore (t‐BPITBT‐TPE) with strong aggregate‐state photoluminescence and good biocompatibility is presented. The NPs comprised of t‐BPITBT‐TPE aggregates encapsulated within 1,2‐distearoyl‐sn‐glycero‐3‐phosphoethanolamine‐N‐[methoxy(polyethylene glycol) (DSPE‐mPEG) micelles exhibit a photoluminescence peak at 660 nm with a high fluorescence quantum yield of 32% in aqueous media. The NPs can be facilely charged by using the same polymeric matrix with different terminal groups, e.g., methoxy (DSPE‐mPEG), amine (DSPE‐PEG‐NH₂), or carboxymethyl (DSPE‐PEG‐COOH) groups. The biocompatibility, toxicity, circulation, and biodistribution of the NPs are assessed using the zebrafish model through whole embryo soaking and intravenous delivery. Furthermore, HeLa and MCF‐7 cells tagged with t‐BPITBT‐TPE in DSPE‐PEG‐NH₂‐TAT polymer NPs are xenografted into zebrafish larvae to successfully track the cancer cell proliferation and metastasis, demonstrating that these new NPs are efficient cancer cell trackers. In addition, the NPs also show good in vivo imaging ability toward 4T1 tumors in xenografted BALB/c mice.     

A Substitution‐Dependent Light‐Up Fluorescence Probe for Selectively Detecting Fe3+ Ions and Its Cell Imaging Application

Deliberate design of specific and sensitive molecular probes with distinctive physical/chemical properties for analyte sensing is of great significance. Herein, by taking advantage of the position‐dependent substituent effects, an aggregation‐induced emission featured iron (III) probe from ortho‐substituted pyridinyl‐functionalized tetraphenylethylene (TPE‐o‐Py) is synthesized. It displays high sensitivity and selectivity toward iron (III) detection. The recognition arises from the position isomer of ortho‐substitution, and the fact that TPE‐o‐Py has a low acid dissociation constant (pK _a) that is close to that of hydrolyzed Fe³⁺. Importantly, TPE‐o‐Py as a light‐up fluorescence probe could be employed for Fe³⁺ sensing in living cells with a pronounced red‐shift in fluorescence emission.     

Precise Molecular Engineering of Photosensitizers with Aggregation‐Induced Emission over 800 nm for Photodynamic Therapy

Owing to efficient singlet oxygen (¹O₂) generation in aggregate state, photosensitizers (PSs) with aggregation‐induced emission (AIE) have attracted much research interests in photodynamic therapy (PDT). In addition to high ¹O₂ generation efficiency, strong molar absorption in long‐wavelength range and near‐infrared (NIR) emission are also highly desirable, but difficult to achieve for AIE PSs since the twisted structures in AIE moieties usually lead to absorption and emission in short‐wavelength range. In this contribution, through acceptor engineering, a new AIE PS of TBT is designed to show aggregation‐induced NIR emission centered at 810 nm, broad absorption in the range between 300 and 700 nm with a large molar absorption coefficient and a high ¹O₂ generation efficiency under white light irradiation. Further, donor engineering by attaching two branched flexible chains to TBT yielded TBTC8 , which circumvented the strong intermolecular interactions of TBT in nanoparticles (NPs), yielding TBTC8 NPs with optimized overall performance in ¹O₂ generation, absorption, and emission. Subsequent PDT results in both in vitro and in vivo studies indicate that TBTC8 NPs are promising candidates in practical application.     

Biocompatible Nanoparticles Based on Diketo‐Pyrrolo‐Pyrrole (DPP) with Aggregation‐Induced Red/NIR Emission for In Vivo Two‐Photon Fluorescence Imaging

Compared with traditional one‐photon fluorescence imaging, two‐photon fluorescence imaging techniques have shown advantages such as increased penetration depth, lower tissue autofluorescence, and reduced photo­damage, and therefore are particularly useful for imaging tissues and animals. In this work, the design and synthesis of two novel DPP ‐based compounds with large two‐photon absorption (2PA) cross‐sections (σ ≥ 8100 GM) and aggregation‐induced emission (AIE) properties are reported. The new compounds are red/NIR emissive and show large Stokes shifts (Δλ ≥ 3571 cm^?1). 1,2‐Distearoyl‐sn‐glycero‐3‐phosphoethanol amine‐N‐[maleimide(polyethylene glycol)‐2000 (DSPE‐PEG‐Mal) is used as the encapsulation matrix to encapsulate DPP‐2 , followed by surface functionalization with cell penetrating peptide (CPP) to yield DPP‐2‐CPP nanoparticles with high brightness, good water dispersibility, and excellent biocompatibility. DPP‐2 nanoparticles have been used for cell imaging and two‐photon imaging with clear visualization of blood vasculature inside mouse ear skin with a depth up to 80 μm.     

Significant Enhancement of Photothermal and Photoacoustic Efficiencies for Semiconducting Polymer Nanoparticles through Simply Molecular Engineering

Semiconducting polymer nanoparticles (SP NPs) are employed as efficient nanoagents for “all‐in‐one” theranostic nanoplatforms with dual photoacoustic imaging (PAI) and photothermal therapy (PTT) functions based on their photothermal conversion effect. However, the mechanisms of tuning the PTT efficiency are still elusive, though several SP NPs with high photothermal efficiency are reported. Herein, two donor–acceptor (D–A) SP NPs PTIGSVS and PIIGSVS with the same donor unit but different acceptor units are designed and synthesized. Through tuning the acceptor unit, PTIGSVS shows more planar backbone structure, stronger D–A strength, redshifted absorption, enhanced extinction efficient, weakened emission properties, and more efficient nonradiative decay in comparison to the polymeric analogue PIIGSVS . Thus, PTIGSVS NPs present much higher photothermal conversion efficiencies (74%) than PIIGSVS NPs (11%), resulting in significantly enhanced in vitro and in vivo PAI and PTT performance. This contribution demonstrates that PTIGSVS NPs are superior PA/PTT agents for effective cancer theranostic and shed light on understanding the relationship between molecular structures and photothermal effect of CPs.     

An NIR‐II Fluorescence/Dual Bioluminescence Multiplexed Imaging for In Vivo Visualizing the Location,Survival, and Differentiation of Transplanted Stem Cells

The in vivo distribution, viability, and differentiation capability of transplanted stem cells are vital for the therapeutic efficacy of stem cell–based therapy. Herein, an NIR‐II fluorescence/dual bioluminescence multiplexed imaging method covering the visible and the second near‐infrared window from 400 to 1700 nm is successfully developed for in vivo monitoring the location, survival, and osteogenic differentiation of transplanted human mesenchymal stem cells (hMSCs) in a calvarial defect mouse model. The exogenous Ag₂S quantum dot–based fluorescence imaging in the second near‐infrared window is applied for visualizing the long‐term biodistribution of transplanted hMSCs. Endogenous red firefly luciferase (RFLuc)‐based bioluminescence imaging (BLI) and the collagen type 1 promoter–driven Gaussia luciferase (GLuc)‐based BLI are employed to report the survival and osteogenic differentiation statuses of the transplanted hMSCs. Meanwhile, by integrating the three imaging channels, multiple dynamic biological behaviors of transplanted hMSCs and the promotion effects of immunosuppression and the bone morphogenetic protein 2 on the survival and osteogenic differentiation of transplanted hMSCs are directly observed. The novel multiplexed imaging method can greatly expand the capability for multifunctional analysis of the fates and therapeutic capabilities of the transplanted stem cells, and aid in the improvement of stem cell–based regeneration therapies and their clinical translation.     

Molecular Engineering of Blue Fluorescent Molecules Based on Silicon End‐Capped Diphenylaminofluorene Derivatives for Efficient Organic Light‐Emitting Materials

Blue fluorescent materials based on silicone end‐capped 2‐diphenylaminofluorene derivatives are synthesized and characterized. These materials are doped into a 2‐methyl‐9,10‐di‐[2‐naphthyl]anthracene host as blue dopant materials in the emitting layer of organic light‐emitting diode devices bearing a structure of ITO/DNTPD (60 nm)/NPB (30 nm)/emitting layer (30 nm)/Alq₃ (20 nm)/LiF (1.0 nm)/Al (200 nm). All devices exhibit highly efficient blue electroluminescence with high external quantum efficiencies (3.47%–7.34% at 20 mA cm^?2). The best luminous efficiency of 11.2 cd A^?1 and highest quantum efficiency of 7.34% at 20 mA cm^?2 are obtained in a device with CIE coordinates (0.15, 0.25). A deep‐blue OLED with CIE coordinates (0.15, 0.14) exhibits a luminous efficiency of 3.70 cd A^?1 and quantum efficiency of 3.47% at 20 mA cm^?2.     

Bio‐Erasable Intermolecular Donor–Acceptor Interaction of Organic Semiconducting Nanoprobes for Activatable NIR‐II Fluorescence Imaging

Activatable second near‐infrared window (NIR‐II; 1.0–1.7 µm) fluorescence probes that uncage deep‐tissue penetrating fluorescence by disease‐related biomarker stimuli hold great promise for detecting diseases with a poor understanding of the pathology at the molecular level with unprecedented resolution. However, currently, very few activatable NIR‐II fluorescence probes are reported mainly due to the lack of a simple yet general design strategy. Herein, a new and fairly generic design strategy using a bio‐erasable intermolecular donor–acceptor interaction to construct activatable NIR‐II fluorescence probes is reported. An organic semiconducting nanoprobe (SPNP) is constructed through blending a biomarker‐sensitive organic semiconducting non‐fullerene acceptor (3,9‐bis(2‐methylene‐(3‐(1,1‐dicyanomethylene)‐cyclopentane‐1,3‐dione‐[c]thiophen))‐5,5,11,11‐tetrakis(4‐hexylphenyl)‐dithieno[2,3‐d:2',3'‐d']‐s‐indaceno[1,2‐b:5,6‐b'] dithiophene) (ITTC) (one of electric acceptors in organic solar cells) with a biomarker‐inert semiconducting polymer donor 5‐(4,8‐bis((2‐ethylhexyl)oxy)‐6‐methylbenzo[1,2‐b:4,5‐b']difuran‐2‐yl)‐10‐methylnaphtho[1,2‐c:5,6‐c']bis([1,2,5]thiadiazole) (PDF) in an amphiphilic‐polymer‐coated single nanoparticle to suppress NIR‐II fluorescence of the donor via a intermolecular donor–acceptor interaction. The acceptor ITTC is found to be specifically degraded by hypochlorite (an important biomarker) to erase its acceptor property, thus erasing the intermolecular donor–acceptor interaction and uncaging NIR‐II fluorescence. Consequently, SPNP exhibits a 17.5‐fold higher fluorescence brightness in the hypochlorite‐abnormal inflammation in vivo than in normal tissues. Our bio‐erasable intermolecular donor–acceptor interaction strategy provides simple yet general guidelines to design various biomarker‐activatable NIR‐II fluorescence probes.     

Molecular Probe Crossing Blood–Brain Barrier for Bimodal Imaging–Guided Photothermal/Photodynamic Therapies of Intracranial Glioblastoma

Currently, treatment of intracranial diseases still remains a great challenge because the blood–brain barrier (BBB) blocks access of most drugs to the central nervous system. Herein, a theranostic small molecular probe, iRGD‐ICG‐Lys‐DTPA@Gd (iRGD‐ILD), capable of crossing BBB is developed. Owing to the small molecular size and α_vβ₃ integrin receptor–mediated transcytosis, this tailor‐made molecular probe integrating the fluorescence and magnetic resonance imaging functions effectively passes through BBB to target tumor cells even in the early stage of glioblastoma multiforme (GBM), thereby allowing a bimodal imaging–guided therapy of GBM. The reactive oxygen species and heat generated by the ICG moiety under the 808 nm laser irradiation exert photodynamic/photothermal therapeutic effects, which results in significantly inhibited tumor growth and prolonged median survival of C6‐Luc glioma‐bearing mice. Notably, the integration of FDA‐approved clinically available agents, e.g., ICG, DTPA and Gd, into a molecular probe may ensure desirable biocompatibility and biosafety for in vivo applications. Overall, the results highlight the potential of a water‐soluble small molecule as a novel theranostic probe for highly effective GBM treatment.