Molecular Oligomerization and Donor Engineering Strategies for Achieving Superior NIR-II Fluorescence Imaging and Thermotherapy under 1064 nm Laser Irradiation

An enormous challenge still exists for designing molecules with the second near-infrared (NIR-II, 1000–1700 nm) window absorption, NIR-II fluorescence emission, and batch-to-batch reproducibility, which is the premise for high-performance NIR-II phototheranostics. Although organic small molecules and polymers have been largely explored for phototheranostics, it is difficult to satisfy the above three elements simultaneously. In this work, molecular oligomerization (the general structure is S-D-A-D′-A-D-S) and donor engineering (changing the donor linker D') strategies are applied to design phototheranostic agents. Such strategies are proved to be efficient in adjusting molecular configuration and energy level, affecting the optical and thermal properties. Three oligomers (O–T, O–DT, and O–Q) are further prepared into water-soluble nanoparticles (NPs). Particularly, the O-T NPs exhibit a higher molar extinction coefficient at 1064 nm (≈4.3-fold of O-DT NPs and ≈4.8-fold of O-Q NPs). Furthermore, the O–T NPs show the highest NIR-II fluorescence brightness and heating capacity (PCE = 73%) among the three NPs under 1064 nm laser irradiation and served as agents for NIR-II imaging guided in vivo photothermal therapy. Overall, by using molecular oligomerization and donor engineering strategies, a powerful example of constructing high-performance NIR-II phototheranostics for clinical translation is given.     

Single Atom-Engineered NIR-II Gold Clusters with Ultrahigh Brightness and Stability for Acute Kidney Injury

Near-infrared-II (NIR-II) imaging has shown great potential for monitoring the pathological progression and deep tissue imaging but is limited to present unmet NIR-II agent. Present fluorophores show a promising prospect for NIR-II imaging, but brightness and photostability are still highly challenging during real-time monitoring. In this work, atom-engineered NIR-II Au₂₄Cd₁ clusters with ultrahigh brightness, stability, and photostability are developed via single atomic Cd doping. Single atom Cd substitutions contribute to Cd 4d state in HOMO and redistribution of energy level near the gap, exhibiting 56-fold fluorescence enhancement of Au₂₄Cd₁ clusters. Meanwhile, single atomic Cd reinforces Cd Au bond energy, formation energy, and stabilized cluster structure, leading to persistent stability for up to 1 month without decay, as well as excellent photostability of 1 h without photobleaching, much longer than clinically approved indocyanine green (<5 min). In vivo imaging shows gold clusters can monitor acute kidney injury (AKI) even after 72 h of injury, enabling evaluating progression at a very long window. Meanwhile, the bioactive gold clusters can alleviate AKI-induced oxidative stress damage and acute neuroinflammation. Single atom-engineered gold clusters exhibit molecular tracking and diagnostic prospect in kidney-related diseases.     

NIR-II Self-Luminous Molecular Probe for In Vivo Inflammation Tracking and Cancer PDT Effect Self-Evaluating

Optical imaging in the second near-infrared (NIR-II, 900–1700 nm) window has been extensively investigated for bioimaging. However, a strong autofluorescence background from real-time excitation light significantly reduces the images’ quality of NIR-II fluorescence (FL) imaging. To resolve this issue, a NIR-II self-luminous small molecule (CLPD) based on bioluminescence (BL) resonance energy transfer (BRET) mechanism is first developed. The reactive oxygen species (ROS) can trigger NIR-II BL and reduce the NIR-II FL signals of the CLPD simultaneously, enabling ROS-correlated ratiometric BL/FL imaging. CLPD is used for high-contrast NIR-II BL imaging of osteoarthritis as well as guiding the treatment process by ratiometric BL/FL imaging. Moreover, CLPD is applied for NIR-II BL imaging of tumor triggered by the generated ROS during PDT. A correlation between the ratiometric NIR-II BL/FL signal and tumor size is constructed, providing a trustworthy tool for early assessment of PDT effect. Overall, this study presents a novel NIR-II self-luminous small molecular probe for in vivo imaging and provides a strategy for design a self-evaluation system of therapeutic effect.     

An Activatable Phototheranostic Nanoplatform for Tumor Specific NIR-II Fluorescence Imaging and Synergistic NIR-II Photothermal-Chemodynamic Therapy

The phototheranostics in the second near-infrared window (NIR-II) have proven to be promising for the precise cancer theranostics. However, the non-responsive and “always on” imaging mode lacks the selectivity, leading to the poor diagnosis specificity. Herein, a tumor microenvironment (TME) activated NIR-II phototheranostic nanoplatform (Ag₂S-Fe(III)-DBZ Pdots, AFD NPs) is designed based on the principle of Förster resonance energy transfer (FRET). The AFD NPs are fabricated through self-assembly of Ag₂S QDs (NIR-II fluorescence probe) and ultra-small semiconductor polymer dots (DBZ Pdots, NIR-II fluorescence quencher) utilizing Fe(III) as coordination nodes. In normal tissues, the AFD NPs maintain in “off” state, due to the FRET between Ag₂S QDs and DBZ Pdots. However, the NIR-II fluorescence signal of AFD NPs can be rapidly “turn on” by the overexpressed GSH in tumor tissues, achieving a superior tumor-to-normal tissue (T/NT) signal ratio. Moreover, the released Pdots and reduced Fe(II) ions provide NIR-II photothermal therapy (PTT) and chemodynamic therapy (CDT), respectively. The GSH depletion and NIR-II PTT effect further aggravate CDT mediated oxidative damage toward tumors, achieving the synergistic anti-tumor therapeutic effect. The work provides a promising strategy for the development of TME activated NIR-II phototheranostic nanoprobes.     

An NIR-II-Emissive Photosensitizer for Hypoxia-Tolerant Photodynamic Theranostics

As a common feature in a majority of malignant tumors, hypoxia has become the Achilles’ heel of photodynamic therapy (PDT). The development of type-I photosensitizers that show hypoxia-tolerant PDT efficiency provides a straightforward way to address this issue. However, type-I PDT materials have rarely been discovered. Herein, a π-conjugated molecule with A–D–A configuration, COi6-4Cl, is reported. The H₂O-dispersible nanoparticle of COi6-4Cl can be activated by an 880 nm laser, and displays hypoxia-tolerant type I/II combined PDT capability, and more notably, a high NIR-II fluorescence with a quantum yield over 5%. Moreover, COi6-4Cl shows a negligible photothermal conversion effect. The non-radiative decay of COi6-4Cl is suppressed in the dispersed and aggregated state due to the restricted molecular vibrations and distinct intermolecular steric hindrance induced by its four bulky side chains. These features make COi6-4Cl a distinguished single-NIR-wavelength-activated phototheranostic material, which performs well in NIR-II fluorescence-guided PDT treatment and shows an enhanced in vivo anti-tumor efficiency over the clinically approved Chlorin e6, by the equal stresses on hypoxia-tolerant anti-tumor therapy and deep-penetration imaging. Therefore, the great potential of COi6-4Cl in precise PDT cancer therapy against hypoxia challenges is demonstrated.     

Size effect of Au seeds on structure of Au–Pt bimetallic nanoparticles

Au–Pt bimetallic nanoparticles (NPs) were synthesized by a seeded growth method. Au NPs with different sizes were obtained by reducing HAuCl₄ with butyllithium, and AuPt bimetallic NPs were synthesized by reducing H₂PtCl₆ with oleylamine using the pre-synthesized Au NPs as seeds. The size of Au seeds was found to be a key factor on the structure of Au–Pt bimetallic NPs. Using big Au NP seeds (8 nm or 12 nm) resulted in the formation of Au–Pt dendritic structures. While relatively small Au NPs (3 nm) were used as seeds, the fast atomic diffusion inside relatively small bimetallic NPs will result in an Au–Pt alloy formation.     

Molecular Engineering of NIR-II/IIb Emitting AIEgen for Multimodal Imaging-Guided Photo-Immunotherapy

In view of the great challenges related to the complexity and heterogeneity of tumors, efficient combination therapy is an ideal strategy for eliminating primary tumors and inhibiting distant tumors. A novel aggregation-induced emission (AIE) phototherapeutic agent called T-TBBTD is developed, which features a donor–acceptor–donor (D–A–D) structure, enhanced twisted molecule conformation, and prolonged second near-infrared window (NIR-II) emission. The multimodal imaging function of the molecule has significance for its treatment time window and excellent photothermal/photodynamic performance for multimode therapy. The precise molecular structure and versatility provide prospects for molecular therapy for anti-tumor applications. Fluorescence imaging in the NIR-II window offers advantages with enhanced spatial resolution, temporal resolution, and penetration depth. The prepared AIE@R837 NPs also have controllable performance for antitumor photo-immunotherapy. Following local photo-irradiation, AIE@R837 NPs generate abundant heat, and ¹O₂ directly kills tumor cells, induces immunogenic cell death (ICD) as a photo-therapeutic effect, and releases R837, which enhances the synergistic effect of antigen presentation and contributes to the long-lasting protective antitumor immunity. A bilateral 4T1 tumor model revealed that this photo-immunotherapy can eliminate primary tumors. More importantly, it has a significant inhibitory effect on distant tumor growth. Therefore, this method can provide a new strategy for tumor therapy.     

NIR-II Imaging-Guided Mitochondrial-Targeting Organic Nanoparticles for Multimodal Synergistic Tumor Therapy

Effectively interfering energy metabolism in tumor cells and simultaneously activating the in vivo immune system to perform immune attacks are meaningful for tumor treatment. However, precisely targeted therapy is still a huge challenge. Herein, a mitochondrial-targeting phototheranostic system, FE-T nanoparticles (FE-T NPs) are developed to damage mitochondria in tumor cells and change the tumor immunosuppressive microenvironment. FE-T NPs are engineered by encapsulating the near-infrared (NIR) absorbed photosensitizer IR-FE-TPP within amphiphilic copolymer DSPE-SS-PEG-COOH for high-performing with simultaneous mitochondrial-targeting, near-infrared II (NIR-II) fluorescence imaging, and synchronous photothermal therapy (PTT) /photodynamic therapy (PDT) /immune therapy (IMT). In tumor treatment, the disulfide in the copolymer can be cleaved by excess intracellular glutathione (GSH) to release IR-FE-TPP and accumulate in mitochondria. After 808 nm irradiation, the mitochondrial localization of FE-T NPs generated reactive oxygen species (ROS), and hyperthermia, leading to mitochondrial dysfunction, photoinductive apoptosis, and immunogenic cell death (ICD). Notably, in situ enhanced PDT/PTT in vivo via mitochondrial-targeting with FE-T NPs boosts highly efficient ICD toward excellent antitumor immune response. FE-T NPs provide an effective mitochondrial-targeting phototheranostic nanoplatform for imaging-guided tumor therapy.     

Low-Dose NIR-II Preclinical Bioimaging Using Liposome-Encapsulated Cyanine Dyes

Fluorescence imaging in the second near-infrared window (NIR-II, 1000–1700 nm) provides a powerful tool for in vivo structural and functional imaging in deep tissue. However, the lack of biocompatible contrast agents with bright NIR-II emission has hindered its application in fundamental research and clinical trials. Herein, a liposome encapsulation strategy for generating ultrabright liposome-cyanine dyes by restricting dyes in the hydrophobic pockets of lipids and inhibiting the aggregation, as corroborated by computational modeling, is reported. Compared with free indocyanine green (ICG, an US Food and Drug Administration-approved cyanine dye), liposome-encapsulated ICG (S-Lipo-ICG) shows a 38.7-fold increase in NIR-II brightness and enables cerebrovascular imaging at only one-tenth dose over a long period (30 min). By adjusting the excitation wavelength, two liposome-encapsulated cyanine dyes (S-Lipo-ICG and S-Lipo-FD1080) enable NIR-II dual-color imaging. Moreover, small tumor nodules (2–5 mm) can be successfully distinguished and removed with S-Lipo-ICG image-guided tumor surgery in rabbit models. This liposome encapsulation maintains the metabolic pathway of ICG, promising for clinical implementation.     

Ag nanoparticles enhanced near-IR emission from Er3+ ions doped glasses

Vitreous materials containing rare-earth (RE) ions and metallic nanoparticles (NPs) attract considerable interest because the presence of the NPs may lead to an intensification of luminescence. In this work, the characteristics of 1.54 μm luminescence for the Er³⁺ ions doped bismuthate glasses containing Ag NPs were studied under 980 nm excitation. The surface plasmon resonance (SPR) band of Ag NPs appears from 500 to 1500 nm. Transmission electron microscopic (TEM) image reveals that the Ag NPs are dispersed homogeneously with the size from 2 to 7 nm. The strength parameters Ω_t(t = 2, 4, 6), spontaneous emission probability (A), radiative lifetime (τ) and stimulated emission section (σ_em) of Er³⁺ ions were calculated by the Judd–Ofelt theory. When the glass contains 0.2 wt% AgCl, the 1.54 μm fluorescence intensity of Er³⁺ reaches a maximum value, which is 7.2 times higher than that of glass without Ag NPs. The Ag NPs embedded glasses show significantly fluorescence enhancement of Er³⁺ ions by local field enhancement from SPR.     

Promoted NIR-II Fluorescence by Heteroatom-Inserted Rigid-Planar Cores for Monitoring Cell Therapy of Acute Lung Injury

Fluorophores with emission in the second near-infrared (NIR-II) window have displayed salient advantages for biomedical applications. However, exploration of new luminogens with high NIR-II fluorescent brightness is still challenging. Herein, based on the “ring-fusion” strategy, a series of heteroatom-inserted rigid-planar cores is proposed to achieve the bathochromic NIR-II fluorophores with aggregation-induced emission (AIE) performance. Interestingly, one of the representative fluorophores, 4,4′-(5,5′-([1,2,5]thiadiazolo[3,4-i]dithieno[2,3-a:3′,2′-c]phenazine-8,12-diyl)bis(4-octylthiophene-5,2-diyl))bis(N,N-diphenylaniline) (TTQiT), enjoys a maximum emission beyond 1100 nm because of the efficiently narrowed energy bandgap by electron-rich sulfur-atom-inserted core, which is verified by theoretical calculation. Taking advantage of the bright NIR-II emission of TTQiT nanoparticles, the desirable in vivo NIR-II imaging with high signal-to-background ratios is successfully performed and a long-term stem cell tracking in the detection of acute lung injury is further realized. Therefore, it is anticipated that this work will provide a promising molecular engineering strategy to enrich the scope of NIR-II fluorophores for catering to diverse demands in biomedical applications.     

A Tumor-Microenvironment-Responsive Lanthanide–Cyanine FRET Sensor for NIR-II Luminescence-Lifetime In Situ Imaging of Hepatocellular Carcinoma

Deep tissue imaging in the second near-infrared (NIR-II) window holds great promise for widespread fundamental research. However, inhomogeneous signal attenuation due to tissue absorption and scattering hampers its application for accurate in vivo biosensing. Here, lifetime-based in situ hepatocellular carcinoma (HCC) detection in NIR-II region is presented using a tumor-microenvironment (peroxynitrite, ONOO⁻)-responsive lanthanide–cyanine Förster resonance energy transfer (FRET) nanosensor. A specially designed ONOO⁻-responsive NIR-II dye, MY-1057, is synthesized as the FRET acceptor. Robust lifetime sensing is demonstrated to be independent of tissue penetration depth. Tumor lesions are accurately distinguished from normal tissue due to the recovery lifetime. Magnetic resonance imaging and liver dissection results illustrate the reliability of lifetime-based detection in single and multiple HCC models. Moreover, the ONOO⁻ amount can be calculated according to the standard curve.     

A mini-review on rare-earth down-conversion nanoparticles for NIR-II imaging of biological systems

Rare-earth (RE) based luminescent probes exhibit rich optical properties including upconversion and down-conversion luminescence spanning a broad spectral range from 300 to 3,000 nm, and have generated great scientific and practical interest from telecommunication to biological imaging. While upconversion nanoparticles have been investigated for decades, down-conversion luminescence of RE-based probes in the second near-infrared (NIR-II, 1,000–1,700 nm) window for in vivo biological imaging with sub-centimeter tissue penetration and micrometer image resolution has come into light only recently. In this review, we present recent progress on RE-based NIR-II probes for in vivo vasculature and molecular imaging with a focus on Er³⁺-based nanoparticles due to the down-conversion luminescence at the long-wavelength end of the NIR-II window (NIR-IIb, 1,500–1,700 nm). Imaging in NIR-IIb is superior to imaging with organic probes such as ICG and IRDye800 in the ~ 800 nm NIR range and the 1,000-1,300 nm short end of NIR-II range, owing to minimized light scattering and autofluorescence background. Doping by cerium and other ions and phase engineering of Er³⁺-based nanoparticles, combined with surface hydrophilic coating optimization can afford ultrabright, biocompatible NIR-IIb probe towards clinical translation for human use. The Nd³⁺-based probes with NIR-II emission at 1,050 and 1,330 nm are also discussed, including Nd³⁺ doped nanocrystals and Nd³⁺-organic ligand complexes. This review also points out future directions for further development of multi-functional RE NIR-II probes for biological imaging.

     

Controlled Synthesis of Ag2Te@Ag2S Core–Shell Quantum Dots with Enhanced and Tunable Fluorescence in the Second Near‐Infrared Window

Fluorescence in the second near‐infrared window (NIR‐II, 900–1700 nm) has drawn great interest for bioimaging, owing to its high tissue penetration depth and high spatiotemporal resolution. NIR‐II fluorophores with high photoluminescence quantum yield (PLQY) and stability along with high biocompatibility are urgently pursued. In this work, a Ag‐rich Ag₂Te quantum dots (QDs) surface with sulfur source is successfully engineered to prepare a larger bandgap of Ag₂S shell to passivate the Ag₂Te core via a facile colloidal route, which greatly enhances the PLQY of Ag₂Te QDs and significantly improves the stability of Ag₂Te QDs. This strategy works well with different sized core Ag₂Te QDs so that the NIR‐II PL can be tuned in a wide range. In vivo imaging using the as‐prepared Ag₂Te@Ag₂S QDs presents much higher spatial resolution images of organs and vascular structures as compared with the same dose of Ag₂Te nanoprobes administrated, suggesting the success of the core–shell synthetic strategy and the potential biomedical applications of core–shell NIR‐II nanoprobes.     

AIE Nanoparticles with High Stimulated Emission Depletion Efficiency and Photobleaching Resistance for Long‐Term Super‐Resolution Bioimaging

Stimulated emission depletion (STED) nanoscopy is a typical super‐resolution imaging technique that has become a powerful tool for visualizing intracellular structures on the nanometer scale. Aggregation‐induced emission (AIE) luminogens are ideal fluorescent agents for bioimaging. Herein, long‐term super‐resolution fluorescence imaging of cancer cells, based on STED nanoscopy assisted by AIE nanoparticles (NPs) is realized. 2,3‐Bis(4‐(phenyl(4‐(1,2,2‐triphenylvinyl)phenyl)amino)phenyl) fumaronitrile (TTF), a typical AIE luminogen, is doped into colloidal mesoporous silica to form fluorescent NPs. TTF@SiO₂ NPs bear three significant features, which are all essential for STED nanoscopy. First, their STED efficiency can reach more than 60%. Second, they are highly resistant to photobleaching, even under long‐term and high‐power STED light irradiation. Third, they have a large Stokes' shift of ≈150 nm, which is beneficial for restraining the fluorescence background induced by the STED light irradiation. STED nanoscopy imaging of TTF@SiO₂‐NPs‐stained HeLa cells is performed, exhibiting a high lateral spatial resolution of 30 nm. More importantly, long‐term (more than half an hour) super‐resolution cell imaging is achieved with low fluorescence loss. Considering that AIE luminogens are widely used for organelle targeting, cellular mapping, and tracing, AIE‐NPs‐based STED nanoscopy holds great potential for many basic biomedical studies that require super‐resolution and long‐term imaging.     

Luminescence Lifetime–Based In Vivo Detection with Responsive Rare Earth–Dye Nanocomposite

For years, luminescence lifetime imaging has served as a quantitative tool in indicating intracellular components and activities. However, very few studies involve the in vivo study of animals, especially in vivo stimuli‐responsive activities of animals, as both excitation and emission wavelengths should fall into the near‐infrared (NIR) optical transparent window (660–950 and 1000–1500 nm). Herein, this work reports a lifetime‐responsive nanocomposite with both excitation and emission in the NIR I window (800 nm) and lifetime in the microsecond region. The incorporation of Tm³⁺‐doped rare‐earth nanocrystals and NIR dye builds an efficient energy transfer pathway that enables a tunable luminescence lifetime range. The NaYF₄:Tm nanocrystal, which absorbs and emits photons at the same energy level, is found to be 33 times brighter than optimized core–shell upconversion nanocrystals, and proved to be an effective donor for NIR luminescence resonance energy transfer (LRET). The anti‐interference capability of luminescence lifetime signals is further confirmed by luminescence and lifetime imaging. In vivo studies also verify the lifetime response upon stimulation generated in an arthritis mouse model. This work introduces an intriguing tool for luminescence lifetime–based sensing in the microsecond region.     

Synthesis and functionalization of SiO2 coated Fe3O4 nanoparticles with amine groups based on self-assembly

The purpose of this research was to synthesize amino modified Fe₃O₄/SiO₂ nanoshells for biomedical applications. Magnetic iron-oxide nanoparticles (NPs) were prepared via co-precipitation. The NPs were then modified with a thin layer of amorphous silica. The particle surface was then terminated with amine groups. The results showed that smaller particles can be synthesized by decreasing the NaOH concentration, which in our case this corresponded to 35 nm using 0.9 M of NaOH at 750 rpm with a specific surface area of 41 m² g^? 1 for uncoated Fe₃O₄ NPs and it increased to about 208 m² g^?1 for 3-aminopropyltriethoxysilane (APTS) coated Fe₃O₄/SiO₂ NPs. The total thickness and the structure of core-shell was measured and studied by transmission electron microscopy (TEM). For uncoated Fe₃O₄ NPs, the results showed an octahedral geometry with saturation magnetization range of (80–100) emu g^?1 and coercivity of (80–120) Oe for particles between (35–96) nm, respectively. The Fe₃O₄/SiO₂ NPs with 50 nm as particle size, demonstrated a magnetization value of 30 emu g^?1. The stable magnetic fluid contained well-dispersed Fe₃O₄/SiO₂/APTS nanoshells which indicated monodispersity and fast magnetic response.     

Electrospun Fe3O4/PVP//Tb(BA)3phen/PVP magnetic–photoluminescent bifunctional bistrand aligned composite nanofibers bundles

Fe₃O₄/PVP//Tb(BA)₃phen/PVP magnetic–photoluminescent bifunctional bistrand aligned composite nanofibers bundles based on Fe₃O₄ nanoparticles (NPs) and terbium complex Tb(BA)₃phen (BA = benzoic acid) were fabricated by employing a parallel axial electrospinning setup and were characterized by X-ray diffraction, field-emission scanning electron microscopy (FESEM), energy dispersive spectroscopy (EDS), transmission electron microscopy, fluorescence spectroscopy, and vibrating sample magnetometer. It is found that Fe₃O₄ NPs were only dispersed into one strand of the bistrand aligned composite nanofibers bundles, but no nanoparticles in the other strand. And the average diameter of the individual strand fiber was 200 ± 25 nm. The bistrand aligned composite nanofibers bundles exhibit strong green emissions under the excitation of 275 nm ultraviolet light, and the ⁵ D ₄ → ⁷ F ₅ hypersensitive transition at 545 nm was the predominant emission peak of Tb³⁺ ions. The newly obtained bifunctional nanofibers bundles exhibit excellent magnetism and high fluorescence intensity and are expected to apply in biology cell separation, magnetic resonance imaging, drug deliver, and fluorescence immunoassays/imaging.     

Bright Far‐Red/Near‐Infrared Conjugated Polymer Nanoparticles for In Vivo Bioimaging

A highly emissive far‐red/near‐infrared (FR/NIR) fluorescent conjugated polymer (CP), poly[(9,9‐dihexylfluorene)‐co‐2,1,3‐benzothiadiazole‐co‐4,7‐di(thiophen‐2‐yl)‐2,1,3‐benzothiadiazole] (PFBTDBT10) is designed and synthesized via Suzuki polymerization. Formulation of PFBTDBT10 using 1,2‐distearoyl‐sn‐glycero‐3‐phosphoethanolamine‐N‐[methoxy(polyethylene glycol)‐2000] (DSPE‐PEG₂₀₀₀) and DSPE‐PEG₅₀₀₀‐folate as the encapsulation matrix yielded CP‐loaded DSPE‐PEG‐folic acid nanoparticles (CPDP‐FA NPs) with bright FR/NIR fluorescence (27% quantum yield) and a large Stoke's shift of 233 nm in aqueous solution. CPDP‐FA NPs show improved thermal/photostabilities and larger Stoke's shifts as compared to commercially available quantum dots (Qdot 655) and organic dyes such as Alexa Fluor 555 and Rhodamine 6G. In vivo studies of CPDP‐FA NPs on a hepatoma H22 tumor‐bearing mouse model reveal that they could serve as an efficient FR/NIR fluorescent probe for targeted in vivo fluorescence imaging and cancer detection in a high contrast and specific manner. Together with the negligible in vivo toxicity, CPDP‐FA NPs are promising FR/NIR fluorescent probes for future in vivo applications.     

Low‐Saturation‐Intensity,High‐Photostability,and High‐Resolution STED Nanoscopy Assisted by CsPbBr3 Quantum Dots

Stimulated emission depletion (STED) nanoscopy is one of the most promising super‐resolution imaging techniques for microstructure imaging. Commercial CdSe@ZnS quantum dots are used as STED probes and ≈50 nm lateral resolution is obtained. Compared with other quantum dots, perovskite CsPbBr₃ nanoparticles (NPs) possess higher photoluminescence quantum yield and larger absorption cross‐section, making them a more effective probe for STED nanoscopy. In this study, CsPbBr₃ NPs are used as probes for STED nanoscopy imaging. The fluorescence intensity of the CsPbBr₃ sample is hardly weakened at all after 200 min irradiation with a 39.8 mW depletion laser, indicating excellent photobleaching resistance of the CsPbBr₃ NPs. The saturation intensity of the CsPbBr₃ NPs is extremely low and estimated to be only 0.4 mW (0.126 MW cm^?2). Finally, an ultrahigh lateral resolution of 20.6 nm is obtained for a single nanoparticle under 27.5 mW STED laser irradiation in CsPbBr₃‐based STED nanoscopy imaging, which is a tenfold improvement compared with confocal microscopy. Because of its high fluorescence stability and ultrahigh resolution under lower depletion power, CsPbBr₃‐assisted STED nanoscopy has great potential to investigate microstructures that require super‐resolution and long‐term imaging.