NIR-II Window Research Articles

Superconducting nanowire single-photon detector enhanced near-infrared II portable confocal microscopy for tissue imaging with indocyanine green.

In this Letter a novel, to our knowledge, approach for near-infrared (NIR) fluorescence portable confocal microscopy is introduced, aiming to enhance fluorescence imaging of biological samples in the NIR-II window. By integrating a superconducting nanowire single-photon detector (SNSPD) into a confocal microscopy, we have significantly leveraged the detection efficiency of the NIR-II fluorescence signal from indocyanine green (ICG), an FDA-approved dye known for its NIR-II fluorescence capabilities. The SNSPD, characterized by its extremely low dark count rate and optimized NIR system detection efficiency, enables the excitation of ICG with 1 mW and the capture of low-light fluorescence signals from deep regions (up to 512 µm). Consequently, our technique was able to produce high-resolution images of bio samples with a superior signal-to-noise ratio, making a substantial advancement in the field of fluorescence microscopy and offering a promising opportunity for future clinical study.

Investigation of the Structure and Optical Properties of Polymethine-Based NIR-II Fluorophores Using Many-Body Perturbation Theory: GW-BSE Approaches.

Fluorescence imaging is a widely used technique for detecting pathophysiological microenvironments and guiding fluorescence-guided therapy owing to its noninvasiveness, high spatiotemporal resolution, ease of operation, and real-time monitoring capabilities. In particular, NIR-II materials are promising for fluorescence imaging applications because they exhibit reduced light scattering and absorption by biological tissues, enabling deeper imaging with improved spatial resolution and contrast compared to visible or first near-infrared imaging. NIR-II materials refer to those that emit in the second near-infrared region of the electromagnetic spectrum, spanning wavelengths from approximately 1000 to 1700 nm. The emission peaks of organic fluorophores within the NIR-II window are of particular interest due to their minimal biotoxicity, in vivo biocompatibility, and biodegradability. In this study, we investigated a new series of NIR-II fluorescent polymethine-based dyes and their NIR-II absorption properties using density functional theory and the GW-BSE approximation. Our calculated maximum absorption peak under the GW-BSE approximation showed good agreement with experimental results, demonstrating the potential of these dyes for NIR-II fluorescence imaging applications.

Scalable Copper Sulfide Formulations for Super-Resolution Optoacoustic Brain Imaging in the Second Near-Infrared Window.

Optoacoustic imaging offers label-free multi-parametric characterization of cerebrovascular morphology and hemodynamics at depths and spatiotemporal resolution unattainable with optical microscopy. Effective imaging depth can greatly be enhanced by employing photons in the second near-infrared (NIR-II) window. However, diminished absorption by hemoglobin along with a lack of suitable contrast agents hinder an efficient application of the technique in this spectral range. Herein, copper sulfide (CuS) micro- and nano-formulations for multi-scale optoacoustic imaging in the NIR-II window are introduced. Dynamic contrast enhancement induced by intravenously administered CuS nanoparticles facilitated visualization of blood perfusion in murine cerebrovascular networks. The individual calcium carbonate microparticles carrying CuS are further shown to generate sufficient responses to enable super-resolution microvascular imaging and blood flow velocity mapping with localization optoacoustic tomography.

Near-Infrared II Fluorescence Imaging Highlights Tumor Angiogenesis in Hepatocellular Carcinoma with a VEGFR-Targeted Probe.

Hepatocellular carcinoma (HCC) is typically characterized by rich vascularity, with angiogenesis playing a crucial role in its growth and invasion. Molecular imaging of specific receptors in blood vessels is crucial in HCC diagnosis. In particular, in vivo imaging utilizing the second near-infrared (NIR-II) window offers improved tissue penetration, reduced light scattering, and lower autofluorescence. Despite the great potential of the NIR-II window, developing safe and effective probes to provide better imaging performance for HCC is urgently needed. In this study, NIR-II imaging integrated with a vascular endothelial growth factor receptor (VEGFR)-targeted probe generated by combining a VEGFR-targeted peptide with indocyanine green (ICG) is used to characterize HCC-related angiogenesis at a resolution of 56.0µm. For the first time, liver metabolic curves and parameters of liver function reserve (LFR) are obtained by fitting NIR-II fluorescence signals with high spatiotemporal resolution, showing significant differences between HCC mice and controls. Moreover, unlike ICG, the targeting probe has a targeted effect on blood vessels in vivo. The tumor-to-normal (T/N) ratio in NIR-II imaging reaches up to 3.30 after post-injection of the targeting probe. The results indicate that the VEGFR-targeted probe is a powerful tool for NIR-II fluorescence imaging to enhance early diagnosis of HCC.

An Integrated Nanoplatform via Dual Channel Excitation for Both Precise Fluorescence Imaging and Photodynamic Therapy of Orthotopic Breast Tumor in NIR-II Region.

Although research on photodynamic therapy (PDT) of malignant tumor has made considerable progress in recent years, it is a remaining challenge to extend PDT to the second near-infrared window (NIR-II) along with real-time and accurate NIR-II fluorescence imaging to determine drug enrichment status and achieve high treatment efficacy. In this work, lanthanide nanoparticles (Ln NPs)-based nanoplatform (LCR) equipped with photosensitizerChlorin e6 (Ce6) and targeting molecular NH2-PEG1000-cRGDfK aredeveloped, which can achieve NIR-II photodynamic therapy (PDT) and NIR-II fluorescence imaging by dual channel excitation. Under 808nm excitation, Nd3+ in the outer layer can absorb the energy and transfer inward to emit strong NIR-II emissions (1064 and 1525nm). Due to the low background noise of NIR-II light and the targeting effect of NH2-PEG1000-cRGDfK, LCR can recognize tiny tumor tissue (≈3mm) and monitor drug distribution in vivo. Under 1530nm excitation, internal Er3+ can be self-sensitized, generating intense upconversion emission (662nm) that can effectively activate Ce6 for in vivo PDT due to the deep tissue penetration of NIR-II light. This study provides a paradigm of theranostic nanoplatform for both real-time fluorescence imaging and PDT of orthotopic breast tumor in NIR-II window.

Croconaine-based NIR-II fluorescence imaging-guided tumor photothermal therapy induces long-term antitumor immune memory

Photothermal therapy (PTT) for cancers guided by optical imaging has recently shown great potential for precise diagnosis and efficient therapy. The second near-infrared window (NIR-II, 1000–1700 nm) fluorescence imaging (FLI) is highly desirable owing to its good spatial and temporal resolution, deep tissue penetration, and negligible tissue toxicity. Organic small molecules are attractive as imaging and treatment agents in biomedical research because of their low toxicity, fast clearance rate, diverse structures, ease of modification, and excellent biocompatibility. Various organic small molecules have been investigated for biomedical applications. However, there are few reports on the use of croconaine dyes (CRs), especially NIR-II emission CRs. To our knowledge, there have been no prior reports of NIR-II emissive small organic photothermal agents (SOPTAs) based on CRs. Herein, we report a croconaine dye (CR-TPE-T)-based nanoparticle (CR NP) with absorption and fluorescence emission in the NIR-I and NIR-II windows, respectively. The CR NPs exhibited intense NIR absorption, outstanding photothermal properties, and good biological compatibility. In vivo studies showed that CR NPs not only achieved real-time, noninvasive NIR-II FLI of tumors, but also induced significant tumor ablation with laser irradiation guided by imaging, without apparent side effects, and promoted the formation of antitumor immune memory in a colorectal cancer model. In addition, the CR NPs displayed efficient inhibition of breast tumor growth, improved longevity of mice and triggered efficient systemic immune responses, which further inhibited tumor metastasis to the lungs. Our study demonstrates the great potential of CRs as therapeutic agents in the NIR-II region for cancer diagnosis.

(Invited) Deep-Brain Imaging and Modulation with Carbon and Polymeric Nanomaterials

Optical approaches to image and modulate brain activity are limited in their penetration depth in live animals, owing to the strong scattering and absorption of photons in the visible spectrum. Carbon nanotubes and carbon-based polymers, in contrast, afford unique optical properties in the second near-infrared window (NIR-II window, 1000-1700 nm) with reduced scattering and absorption by the brain. In this talk, I will present my lab’s latest work on the imaging and modulation of brain activity with carbon nanotubes and carbon-based polymeric nanoparticles in the live mouse brain. Specifically, the NIR-II window enables through-scalp imaging of cerebrovasculature and modulation of deep-brain neural activity in freely behaving animals via a minimally invasive, implant-free, and tether-free optical interface. Our development of carbon- and polymer-based deep-brain imaging and neuromodulation has been published in Nature Photonics (2014), Nature Biomedical Engineering (2017), Nature Biomedical Engineering (2022), and ACS Nano (2023).

Single Molecule Imaging of Sp3 Functionalized Ultrashort Carbon Nanotubes for Deep Brain Tissue Investigation

Near-infrared (NIR) fluorescence microscopy has attracted significant attention for in vivo deep tissue imaging due to the combination of low light scattering and absorption by the tissues, resulting in good light penetration depth, especially in the NIR-II window (~1000–1350 nm). In this context, NIR photoluminescence (PL) (λem ~ 900–1400 nm) of single-walled carbon nanotubes overlapping with the so-called “tissue transparency window” laid the foundation for their use in bioimaging and sensing applications. Interestingly, although ultrashort carbon nanotubes (usCNTs) had long been sought for their advantages of mimicking bio-molecular dimensions, it is only when sp3 defects introduced bright fluorescent usCNTs could be generated [1]. In this work, we demonstrate the efficient preparation of bright fluorescent usCNTs specifically designed for biological applications, and we show how to create bright usCNTs wrapped by a biocompatible encapsulating agent (phospholipid-polyethylene glycol, PL-PEG). More precisely, we adapted a synthesis route where the sp3 functionalization is followed by chemical cutting [2]. We will present their full characterization and demonstrate their performance for deep tissue imaging in neuroscience. More precisely, our results performed on live brain tissue allow us to analyze their length specific behaviors in different compartments of mouse brains in comparison to the long sp3 functionalized CNTs [3].

Extending the emission peak tail of indole cyanine for deep-near-infrared bioimaging

We propose a novel strategy for tailoring the structure of fluorescent molecules to achieve emission at the tail end of the NIR-II window. The favorable spectroscopic properties and low cytotoxicity of YNs make them powerful tools for bioimaging. Notably, YN-4 exhibits a brightness 2.5 times greater than YN-3, 6 times that of IR-783, and 5 times that of ICG. This enhanced brightness enabled high-resolution imaging of mouse thoracic and abdominal cavities, tumor vasculature, and real-time monitoring of gastrointestinal motility using YN-4. Furthermore, covalent grafting of glucose onto the YN-Glu scaffold significantly improved tumor-targeting capability and facilitated tracking of glucose metabolism. This work aims to extend the application of fluorescent molecule imaging beyond the NIR-IIa window.

Tuning the photophysical properties of cyanine by barbiturate functionalization and nanoformulation for efficient optoacoustics- guided phototherapy

Cyanine derivatives are organic dyes widely used for optical imaging. However, their potential in longitudinal optoacoustic imaging and photothermal therapy remains limited due to challenges such as poor chemical stability, poor photostability, and low photothermal conversion. In this study, we present a new structural modification for cyanine dyes by introducing a strongly electron-withdrawing group (barbiturate), resulting in a new series of barbiturate-cyanine dyes (BC810, BC885, and BC1010) with suppressed fluorescence and enhanced stability. Furthermore, the introduction of BC1010 into block copolymers (PEG114-b-PCL60) induces aggregation-caused quenching, further boosting the photothermal performance. The photophysical properties of nanoparticles (BC1010-NPs) include their remarkably broad absorption range from 900 to 1200 nm for optoacoustic imaging, allowing imaging applications in NIR-I and NIR-II windows. The combined effect of these strategies, including improved photostability, enhanced nonradiative relaxation, and aggregation-caused quenching, enables the detection of optoacoustic signals with high sensitivity and effective photothermal treatment of in vivo tumor models when BC1010-NPs are administered before irradiation with a 1064 nm laser. This research introduces a barbiturate-functionalized cyanine derivative with optimal properties for efficient optoacoustics-guided theranostic applications. This new compound holds significant potential for biomedical use, facilitating advancements in optoacoustic-guided diagnostic and therapeutic approaches.

Gold Nanoclusters as High Resolution NIR-II Theranostic Agents.

In the realm of nanomaterials, atomically precise quasi-molecular gold nanoclusters (AuNCs) play a prime role due to their unique, stable, and highly tunable optical properties. They are extensively structure-engineered for modulation of surface electronic states toward long wavelength photoluminescence, particularly in the NIR-II (1000 to 1700 nm) window. Contrast agents with NIR-II emission can potentially transform optical imaging in terms of higher spatial resolution, deeper tissue penetration, and reduced tissue autofluorescence. These advantages allow real-time imaging in living organisms for observing disease progression and treatment response. In this short review, we discuss origin of NIR-II emission in rationally designed AuNCs and their application toward high resolution imaging of vasculatures and hard and soft tissue structures for identification of pathological conditions such as stroke and injury. Further, recent employment of these AuNCs in the rapidly growing field of tumor theranostics is also summarized. Final remarks are provided on the scope for improvement in their optical properties and persisting challenges for clinical translation.

Tumor-specific enhanced NIR-II photoacoustic imaging via photothermal and low-pH coactivated AuNR@PNIPAM-VAA nanogel

BackgroundProperly designed second near-infrared (NIR-II) nanoplatform that is responsive tumor microenvironment can intelligently distinguish between normal and cancerous tissues to achieve better targeting efficiency. Conventional photoacoustic nanoprobes are always “on”, and tumor microenvironment-responsive nanoprobe can minimize the influence of endogenous chromophore background signals. Therefore, the development of nanoprobe that can respond to internal tumor microenvironment and external stimulus shows great application potential for the photoacoustic diagnosis of tumor.ResultsIn this work, a low-pH-triggered thermal-responsive volume phase transition nanogel gold nanorod@poly(n-isopropylacrylamide)-vinyl acetic acid (AuNR@PNIPAM-VAA) was constructed for photoacoustic detection of tumor. Via an external near-infrared photothermal switch, the absorption of AuNR@PNIPAM-VAA nanogel in the tumor microenvironment can be dynamically regulated, so that AuNR@PNIPAM-VAA nanogel produces switchable photoacoustic signals in the NIR-II window for tumor-specific enhanced photoacoustic imaging. In vitro results show that at pH 5.8, the absorption and photoacoustic signal amplitude of AuNR@PNIPAM-VAA nanogel in NIR-II increases up obviously after photothermal modulating, while they remain slightly change at pH 7.4. Quantitative calculation presents that photoacoustic signal amplitude of AuNR@PNIPAM-VAA nanogel at 1064 nm has ~ 1.6 folds enhancement as temperature increases from 37.5 °C to 45 °C in simulative tumor microenvironment. In vivo results show that the prepared AuNR@PNIPAM-VAA nanogel can achieve enhanced NIR-II photoacoustic imaging for selective tumor detection through dynamically responding to thermal field, which can be precisely controlled by external light.ConclusionsThis work will offer a viable strategy for the tumor-specific photoacoustic imaging using NIR light to regulate the thermal field and target the low pH tumor microenvironment, which is expected to realize accurate and dynamic monitoring of tumor diagnosis and treatment.

NIR-II Surface-Enhanced Raman Scattering Nanoprobes in Biomedicine: Current Impact and Future Directions.

The field of second near-infrared (NIR-II) surface-enhanced Raman scattering (SERS) nanoprobes has made commendable progress in biomedicine. This article reviews recent advances and future development of NIR-II SERS nanoprobes. It introduces the fundamental principles of SERS nanoprobes and highlights key advances in the NIR-II window, including reduced tissue attenuation, deep penetration, maximized allowable exposure, and improved photostability. The discussion of future directions includes the refinement of nanoprobe substrates, emphasizing the tailoring of optical properties of metallic SERS-active nanoprobes, and exploring non-metallic alternatives. The intricacies of designing Raman reporters for the NIR-II resonance and the potential of these reporters to advance the field are also discussed. The integration of artificial intelligence (AI) into nanoprobe design represents a cutting-edge approach to overcome current challenges. This article also examines the emergence of deep Raman techniques for through-tissue SERS detection, toward NIR-II SERS tomography. It acknowledges instrumental advancements like improved charge-coupled devicesensitivity and accelerated imaging speeds. The article concludes by addressing the critical aspects of biosafety, ease of functionalization, compatibility, and the path to clinical translation. With a comprehensive overview of current achievements and future prospects, this review aims to illuminate the path for NIR-II SERS nanoprobes to innovate diagnostic and therapeutic approaches in biomedicine.

NIR-II Fluorescent Probes for Fluorescence-Imaging-Guided Tumor Surgery.

Second near-infrared (NIR-II) fluorescence imaging is the most advanced imaging fidelity method with extraordinary penetration depth, signal-to-background ratio, biocompatibility, and targeting ability. It is currently booming in the medical realm to diagnose tumors and is being widely applied for fluorescence-imaging-guided tumor surgery. To efficiently execute this modern imaging modality, scientists have designed various probes capable of showing fluorescence in the NIR-II window. Here, we update the state-of-the-art NIR-II fluorescent probes in the most recent literature, including indocyanine green, NIR-II emissive cyanine dyes, BODIPY probes, aggregation-induced emission fluorophores, conjugated polymers, donor-acceptor-donor dyes, carbon nanotubes, and quantum dots for imaging-guided tumor surgery. Furthermore, we point out that the new materials with fluorescence in NIR-III and higher wavelength range to further optimize the imaging results in the medical realm are a new challenge for the scientific world. In general, we hope this review will serve as a handbook for researchers and students who have an interest in developing and applying fluorescent probes for NIR-II fluorescence-imaging-guided surgery and that it will expedite the clinical translation of the probes from bench to bedside.

Semiconducting Polymers Based on Asymmetric Thiadiazoloquinoxaline for Augmented In Vivo NIR-II Photoacoustic Imaging.

A photoacoustic (PA) imaging technique using the second near-infrared (NIR-II) window has attracted more and more attention because of its merits of deeper penetration depth and higher signal-to-noise (S/N) ratio than that using the first near-infrared (NIR-I) one. However, the design and development of high-performance PA imaging contrast agents in the NIR-II window is still a challenge. A semiconducting polymer, constructed by asymmetric units, exhibits regiorandom characteristics that effectively increase the distortion of the backbone. This increase in the degree of twist can regulate the twisted intramolecular charge transfer (TICT) effect, resulting in an enhancement of the PA signal. In this paper, an asymmetric structural acceptor strategy is developed to improve the PA signals of the resulting semiconducting polymer (PATQ-MP) in the NIR-II window with improved brightness, higher S/N ratio, and better photothermal conversion efficiency compared to polymers with the same main-chain structure containing a symmetric acceptor. DFT analysis showed that PATQ-MP containing an asymmetric acceptor monomer had a larger dihedral angle, which effectively improved the PA signal intensity by enhancing the TICT effect. The PEG-encapsulated PATQ-MP nanoparticles exhibit promising performance in the PA imaging of mouse tumors in vivo, demonstrating the clear identification of microvessels as small as 100 μm along with rapid metabolism within a span of 5 h. Therefore, this work provides a unique molecular design strategy for improving the signal intensity of PA imaging in the NIR-II window.

Samarium doped carbon dots for near-infrared photo-therapy

Photo-assisted tumor therapy, encompassing photodynamic therapy (PDT) and photothermal therapy (PTT), has garnered considerable attention as a promising approach for tumor treatment owing to its stimuli-responsive nature and noninvasive characteristics. Carbon dots (CDs) have recently emerged as a novel type of nanomaterial with great potential in the field of anticancer therapy. However, the development of multifunctional CDs nano-platforms that integrate multiple therapeutic performance remains a challenge, especially in near-infrared (NIR) window. In this study, we synthesized samarium-doped carbon dots (Sm/CDs) via a one-step hydrothermal process for fluorescence imaging guided PTT/PDT in NIR-II window. Sm/CDs demonstrated remarkable absorption and emission properties in the NIR region, enabling efficient generation of heat energy, superoxide (•O2–) and hydroxyl radicals (•OH) under NIR irradiation. The photothermal conversion efficiencies of Sm/CDs reached up to 34.12 % and 24.38 % when exposed to 808 nm and 1060 nm wavelengths, respectively. Moreover, long-term toxicity experiments conducted in vivo as well as fluorescence experiments confirmed the potential of such functional CDs for fluorescence imaging and excellent biocompatibility. It is noteworthy that the tumor is nearly eradicated when exposed to either 808 nm or 1060 nm, demonstrating significant efficacy in both scenarios. This study provides a multifunctional CDs nano-platform that offers a promising bimodal phototherapeutic strategy to enhance the synergistic treatment of tumors through PDT and PTT.

Molecular Engineering of Activatable NIR-II Hemicyanine Reporters for Early Diagnosis and Prognostic Assessment of Inflammatory Bowel Disease.

Molecular imaging in the second near-infrared window (NIR-II) provides high-fidelity visualization of biopathological events in deep tissue. However, most NIR-II probes produce "always-on" output and demonstrate poor signal specificity toward biomarkers. Herein, we report a series of hemicyanine reporters (HBCs) with tunable emission to NIR-II window (715-1188 nm) and structurally amenable to constructing activatable probes. Such manipulation of emission wavelengths relies on rational molecular engineering by integrating benz[c,d]indolium, benzo[b]xanthonium, and thiophene moieties to a conventional hemicyanine skeleton. In particular, HBC4 and HBC5 possess bright and record long emission over 1050 nm, enabling improved tissue penetration depth and superior signal to background ratio for intestinal tract mapping than NIR-I fluorophore HC1. An activatable inflammatory reporter (AIR-PE) is further constructed for pH-triggered site-specific release in colon. Due to minimized background interference, oral gavage of AIR-PE allows clear delineation of irritated intestines and assessment of therapeutic responses in a mouse model of inflammatory bowel disease (IBD) through real-time NIRF-II imaging. Benefiting from its high fecal clearance efficiency (>90%), AIR-PE can also detect IBD and evaluate the effectiveness of colitis treatments via in vitro optical fecalysis, which outperforms typical clinical assays including fecal occult blood testing and histological examination. This study thus presents NIR-II molecular scaffolds that are not only applicable to developing versatile activatable probes for early diagnosis and prognostic monitoring of deeply seated diseases but also hold promise for future clinical translations.

A Nanographene-Porphyrin Hybrid for Near-Infrared-Ii Phototheranostics.

Photoacoustic imaging (PAI)-guided photothermal therapy (PTT) in the second near-infrared (NIR-II, 1000-1700nm) window has been attracting attention as a promising cancer theranostic platform. Here, it is reported that the π-extended porphyrins fused with one or two nanographene units (NGP-1 and NGP-2) can serve as a new class of NIR-responsive organic agents, displaying absorption extending to ≈1000 and ≈1400nm in the NIR-I and NIR-II windows, respectively. NGP-1 and NGP-2 are dispersed in water through encapsulation into self-assembled nanoparticles (NPs), achieving high photothermal conversion efficiency of 60% and 69%, respectively, under 808 and 1064nm laser irradiation. Moreover, the NIR-II-active NGP-2-NPs demonstrated promising photoacoustic responses, along with high photostability and biocompatibility, enabling PAI and efficient NIR-II PTT of cancer in vivo.

Optimum Dimensions for Au Nanoframe Tuning in the Near-Infrared Second Window Range for Biomedical Applications: A Numerical Study

AbstractAu nanoparticles are favored in biomedical applications owing to their low cost and negligible cytotoxicity to biological cells. Nanoframes outshine their solid counterparts because of their porosity, which produces pronounced redshifts in their local surface plasmon resonance (LSPR). This feature enables the utilization of nanoframes in photothermal-based therapy, where LSPR excitation of particles within the near-infrared range (NIR) is essential. LSPR redshift in nanoframes is highly sensitive to their dimensions. A slight difference in the nanoframe dimension can result in substantial redshift, potentially pushing its LSPR beyond or below the required NIR range. We perform a systematic numerical study to investigate the optimum dimensions within a range of 1–100 nm for a spherical frame (SpF) and standard cubic frame (CF) to precisely tune their LSPR within the NIR-II window (1000–1400 nm). Our findings indicate that SpF exhibits a shorter LSPR redshift than CF’s at a certain porosity limit that is related to the geometry of the frame. Moreover, SpF displays higher LSPR sensitivity in the NIR region compared to CF. These insights provide valuable guidance for nanoframe design tailored for photothermal-based biomedical applications.

Applications of nanotheranostics in the second near-infrared window in bioimaging and cancer treatment.

Achieving accurate and efficient tumor imaging is crucial in the field of tumor treatment, as it facilitates early detection and precise localization of tumor tissues, thereby informing therapeutic strategies and surgical interventions. The optical imaging technology within the second near-infrared (NIR-II) window has garnered significant interest for its remarkable benefits, such as enhanced tissue penetration depth, superior signal-to-background ratio (SBR), minimal tissue autofluorescence, reduced photon attenuation, and lower tissue scattering. This review explained the design and optimization strategies of nano-agents responsive to the NIR-II window, such as single-walled carbon nanotubes, quantum dots, lanthanum-based nanomaterials, and noble metal nanomaterials. These nano-agents enable non-invasive, deep-tissue imaging with high spatial resolution in the NIR-II window, and their superior optical properties significantly improve the accuracy, efficiency, and versatility of imaging-guided tumor treatments. And we discussed the characteristics and advantages of fluorescence imaging (FL)/photoacoustic imaging (PA) in NIR-II window, providing a comprehensive overview of the latest research progress of different nano-agents in FL/PA imaging-guided tumor therapy. Furthermore, we exhaustively reviewed the latest applications of multifunctional nano-phototherapy technologies carried out by NIR-II light including photothermal therapy (PTT), photodynamic therapy (PDT), and combined modalities like photothermal-chemodynamic therapy (PTT-CDT), photothermal-chemotherapy (PTT-CT), and photothermal- immunotherapy (PTT-IO). These imaging-guided integrated tumor therapy approaches within the NIR-II window have gradually matured over the past decade and are expected to become a safe and effective non-invasive tumor treatment. Finally, we outlined the prospects and challenges of development and innovation of the NIR-II integrated diagnosis and therapy nanoplatform. This review aims to provide insightful perspectives for future advancements in NIR-II optical tumor diagnosis and integrated treatment platforms.