Nanoprobes For Imaging Research Articles

Rapid Detection of Ag(I) via Size-Induced Photoluminescence Quenching of Biocompatible Green-Emitting, l-Tryptophan-Scaffolded Copper Nanoclusters.

Atomically precise metal nanoclusters capped with small molecules like amino acids are highly favored due to their specific interactions and easy incorporation into biological systems. However, they are rarely explored due to the challenge of surface functionalization of nanoclusters with small molecules. Herein, we report the synthesis of a green-emitting (λex = 380 nm, λem = 500 nm), single-amino-acid (l-tryptophan)-scaffolded copper nanocluster (Trp-Cu NC) via a one-pot route under mild reaction conditions. The synthesized nanocluster can be used for the rapid detection of a heavy metal, silver (Ag(I)), in the nanomolar concentration range in real environmental and biological samples. The strong green photoluminescence intensity of the nanocluster quenched significantly upon the addition of Ag(I) due to the formation of bigger nanoparticles, thereby losing its energy quantization. A notable color change from light yellow to reddish-brown can also be observed in the presence of Ag(I), allowing its visual colorimetric detection. Portable paper strips fabricated with the Trp-Cu NC can be reliably used for on-site visual detection of Ag(I) in the micromolar concentration range. The Trp-Cu NC possesses excellent biocompatibility, making it a suitable nanoprobe for cell imaging; thus, it can act as an in vivo biomarker. The nanocluster showed a significant spectral overlap with anticancer drug doxorubicin and thus can be used as an effective fluorescence resonance energy transfer (FRET) pair. FRET results can reveal important information regarding the attachment of the drug to the nanocluster and hence its role as a potential drug carrier for targeted drug delivery within the human body.

Nontoxic Fluorescent Nanoprobes for Multiplexed Detection and 3D Imaging of Tumor Markers in Breast Cancer

Multiplexed fluorescent immunohistochemical analysis of breast cancer (BC) markers and high-resolution 3D immunofluorescence imaging of the tumor and its microenvironment not only facilitate making the disease prognosis and selecting effective anticancer therapy (including photodynamic therapy), but also provides information on signaling and metabolic mechanisms of carcinogenesis and helps in the search for new therapeutic targets and drugs. The characteristics of imaging nanoprobe efficiency, such as sensitivity, target affinity, depth of tissue penetration, and photostability, are determined by the properties of their components, fluorophores and capture molecules, and by the method of their conjugation. Regarding individual nanoprobe components, fluorescent nanocrystals (NCs) are widely used for optical imaging in vitro and in vivo, and single-domain antibodies (sdAbs) are well established as highly specific capture molecules in diagnostic and therapeutic applications. Moreover, the technologies of obtaining functionally active sdAb-NC conjugates with the highest possible avidity, with all sdAb molecules bound to the NC in a strictly oriented manner, provide 3D-imaging nanoprobes with strong comparative advantages. This review is aimed at highlighting the importance of an integrated approach to BC diagnosis, including the detection of biomarkers of the tumor and its microenvironment, as well as the need for their quantitative profiling and imaging of their mutual location, using advanced approaches to 3D detection in thick tissue sections. The existing approaches to 3D imaging of tumors and their microenvironment using fluorescent NCs are described, and the main comparative advantages and disadvantages of nontoxic fluorescent sdAb-NC conjugates as nanoprobes for multiplexed detection and 3D imaging of BC markers are discussed.

Structural and Molecular Fusion MRI Nanoprobe for Differential Diagnosis of Malignant Tumors and Follow-Up Chemodynamic Therapy.

Enhanced imaging techniques using contrast agents enable high-resolution structural imaging to reveal space-occupying lesions but rarely provide detailed molecular information. To this end, we report a structural and molecular fusion magnetic resonance imaging (MRI) nanoprobe for differential diagnosis between benign and malignant tumors. This fusion nanoprobe, termed FFT NPs, follows a working mechanism involving a T1-/T2-weighted magnetic resonance tuning effect (MRET) between a magnetic Fe3O4 core and a paramagnetic Fe-tannic acid (Fe-TA) shell. The FFT NPs with an "always-on" inert T2 signal provide structural MRI (sMRI) contrast of tumors while affording an activated T1 signal in the presence of ATP, which is overproduced during the rapid growth of malignant tumors to enable molecular MRI (mMRI) of tumor lesions. We propose the use of the ratiometric mMRI:sMRI intensity to assist in the differential diagnosis of malignant 4T1 tumors from benign L929 fibroblast tumors. Furthermore, the dissociated FFT NPs were found to be able to catalyze H2O2 conversion in 4T1 tumors to generate excess reactive oxygen species (ROS) for chemodynamic therapy. The described fusion nanoprobe strategy enables the differential diagnosis of tumors from a combined spatial and molecular perspective with one-stop MRI imaging with potential applications in precision intervention.

Bioorthogonal surface-enhanced Raman scattering flower-like nanoprobe with embedded standards for accurate cancer cell imaging

Developing precise and effective strategies for cancer identification and imaging is attractive due to their importance for early cancer detection, prognosis, and subsequent treatment. Herein, we reported a novel bioorthogonal surface-enhanced Raman scattering (SERS) nanoprobe for accurate cancer cell imaging. A novel core-molecule-shell nanoflower (Au@4-MBN@Au) with rich electromagnetic hot spots and enhanced Raman scattering was first synthesized by optimizing the embedded concentrations of 4-mercaptobenzonitrile (4-MBN). Then, Au@4-MBN@Au was further modified with FA-PEG-SH molecules to acquire the bioorthogonal SERS nanoprobe Au@4-MBN@Au-PEG-FA. The SERS nanoprobe illustrated a robust and stable nitrile stretching vibration Raman signal (2223 cm−1) in the cellular silent region, ensuring high sensitivity and ultra-accuracy SERS imaging of cancer cells. Furthermore, cell imaging results demonstrated Au@4-MBN@Au-PEG-FA could recognize FR-positive HeLa cells with high selectivity due to the high affinity between folate receptor and folic acid. More notably, Au@4-MBN@Au-PEG-FA has been applied to identify FR-positive Hela cells from co-cultured cancer cells with similar morphology by SERS imaging for the first time. With improved signal-to-background ratio, high selectivity, and excellent stability, we anticipate the SERS nanoprobe Au@4-MBN@Au-PEG-FA could be applied for FR-related cancer theranostics and clinical detection in the future.

Fluorophore-Decorated Mie Resonant Silicon Nanosphere for Scattering/Fluorescence Dual-Mode Imaging.

Inorganic nanoparticles with multiple functions have been attracting attention as multimodal nanoprobes in bioimaging, biomolecule detection, and medical diagnosis and treatment. A drawback of conventional metallic nanoparticle-based nanoprobes is the Ohmic losses that lead to fluorescence quenching of attached molecules and local heating under light irradiation. Here, metal-free nanoprobes capable of scattering/fluorescence dual-mode imaging are developed. The nanoprobes are composed of a silicon nanosphere core having efficient Mie scattering in the visible to near infrared range and a fluorophore doped silica shell. The dark-field scattering and photoluminescence images/spectra for nanoprobes made from different size silicon nanospheres and different kinds of fluorophores are studied by single particle spectroscopy. The fluorescence spectra are strongly modified by the Mie modes of a silicon nanosphere core. By comparing scattering and fluorescence spectra and calculated Purcell factors, the fluorescence enhancement factor is quantitatively discussed. In vitro scattering/fluorescence imaging studies on human cancer cells demonstrate that the developed nanoparticles work as scattering/fluorescence dual-mode imaging nanoprobes.

A robust high selectivity fluorescence turn-on nanoprobe for peroxynitrite detection in inflammatory cells and mice

Changed levels of intracellular peroxynitrite anion (ONOO−) are closely related to the occurrence and development of inflammation. Specific imaging of ONOO− at sites of inflammation can be of great significance not only for inflammation diagnosis but also for obtaining a deeper understanding of the role of ONOO− in inflammation. Therefore, there is an urgent need for constructing some reliable tools to study the relationship between ONOO− and inflammation in biosystems. In this work, we developed a robust high selectivity fluorescence turn-on nanoprobe (Rhb-ONOO) for inflammation-targeted imaging of ONOO−. The Rhb-ONOO was obtained by self-assembly of amphiphilic Rhb-ONOO, which was constructed by the condensation reaction of the hydrophobic, ONOO−-response and deep red-emitting fluorophore (Rhb) with hydrophilic biopolymer glycol chitosan (GC). Rhb-ONOO showed rapid response towards ONOO− during 60 s, high sensitivity with 19-fold enhancement of fluorescence intensity ratio (I628/I0), and excellent selectivity towards ONOO− over other analytes as well as a good linear relationship was observed between the I628/I0 and the ONOO− concentration range 0–1 μM, with an excellent limit of detection (LOD) of 33 nM. Impressively, it was successfully employed Rhb-ONOO for ONOO− imaging in living inflammatory cells and drug-induced inflammatory mice, illustrating nanoprobe Rhb-ONOO has excellent potential for further study ONOO−-related inflammatory diseases.

Preparations of antibacterial yellow-green-fluorescent carbon dots and carbon dots-lysozyme complex and their applications in bacterial imaging and bacteria/biofilm inhibition/clearance

The preparation of functional long-wavelength-emitting nanomaterials and the researches on their applications in antibacterial and antibiofilm fields have important significance. This paper reports the preparation of yellow-green-fluorescent and high- quantum yield carbon dots (4-ACDs) with 4-aminosalicylic acid and polyethylene imine as raw materials through one-step route, and the impacts of raw material structure and the reaction conditions upon the optical properties of the products have been investigated. 4-ACDs exhibit excellent broad-spectrum antibacterial activity, and their good biocompatibility ensures them as ideal fluorescent nano-probe for cell imaging. However, 4-ACDs could not effectively eliminate the biofilm of Staphylococcus aureus (S. aureus). CDs-LZM complex was prepared through the coupling between 4-ACDs and lysozyme (LZM) and the complex showed strong antibacterial activity against Gram-positive bacteria, particularly with MIC against S. aureus at 5 μg mL−1. Besides, CDs-LZM showed excellent ability against the biofilm of S. aureus. At the concentration of 60 μg mL−1, its inhibition rate against the growth of biofilm was 86 %, and elimination rate against biofilm reached 76 %. CDs-LZM exhibited obvious antibiofilm ability through removing extracellular matrix of biofilm, greatly reducing the thickness of biofilm under confocal microscopy. The application of novel long-wavelength-emitting nanomaterial in eliminating pathogenic bacteria is of great significance.

Multifunctional nanoprobe for multi-mode imaging and diagnosis of metastatic prostate cancer

The high incidence and complex subtypes of prostate cancer put forward higher requirements for accurate diagnosis. Furthermore, advanced prostate cancer is prone to metastasis. Single biological imaging mode faces a challenge of sensitive and fast bioimaging of metastasic prostate cancer. Thus, exploring a nanoprobe with multi-mode imaging function has an important impact on preoperative imaging and intraoperative visualization guide of metastatic prostate cancer. Herein, based on the optical properties and X-ray attenuation capability of Au nanodots as well as the slow electronic relaxation of Gd3+, we designed and fabricated the multifunctional nanoprobe Au/Gd nanodots for multi-mode imaging and accurate diagnosis of bone metastatic prostate cancer. The results showed that multiple imaging modes complement each other to achieve high-precision of metastasic prostate cancer detection and accurately guide treatment. In addition, in vitro/vivo experiments showed that Au/Gd nanodots had good biocompatibility and biosafety. Therefore, the prepared multifunctional nanoprobe may provide new strategies and insights for precise diagnosis of metastatic prostate cancer in clinical practice.

Near-Infrared Fluorescent Nanoprobes for Adenosine Triphosphate-Guided Imaging in Cancer and Fatty Liver Mice.

Adenosine triphosphate (ATP), as an indispensable biomolecule, is the main energy source of cells and is used as a marker for diseases such as cancer and fatty liver. It is of great significance to design a near-infrared fluorescent nanoprobe with excellent performance and apply it to various disease models. Here, a near-infrared fluorescent nanoprobe (ZIF-90@SiR) based on a zeolitic imidazole framework is proposed. The fluorescent nanoprobes are synthesized by encapsulating the dye (SiR) into the framework of ZIF-90. Upon the addition of ATP, the structure of the ZIF-90@SiR nanoprobe is disrupted and SiR is released to generate near-infrared fluorescence at 670 nm. In the process of ATP detection, ZIF-90@SiR shows high sensitivity and good selectivity. Moreover, the ZIF-90@SiR nanoprobe has good biocompatibility due to its low toxicity to cells. It is used for fluorescence imaging of ATP in living cells and thus distinguishing normal cells and cancer cells, as well as distinguishing fatty liver cells. Due to excellent near-infrared fluorescence properties, the ZIF-90@SiR nanoprobe can not only distinguish normal mice and tumor mice but also differentiate normal mice and fatty liver mice for the first time.

Constructing bimodal nanoprobe based on Gd:AgInS2/ZnS quantum dots for fluorometric/magnetic resonance imaging in mesenchymal stem cells

Mesenchymal stem cells (MSCs) hold great promise in regenerative medicine and received overwhelming concerns to promote their therapeutic effects. Owing to the shortage of MSCs-specific biomarkers, bimodal imaging nanoprobes with integrated complementary information are of great importance in ameliorating the efficacy of MSCs terminal tracking. In this study, a noninvasive dual-mode imaging nanoprobe with enhanced detection sensitivity and spatial resolution based on alloyed Gd:AgInS2/ZnS quantum dots (QDs) was first fabricated through a microwave-assisted heating method. The QDs with red emissive fluorescence exhibit excellent biocompatibility in MSCs under a confocal microscope. As for magnetic resonance imaging (MRI), the longitudinal relaxation rate of 11.1420 mM–1 S–1 of Gd:AgInS2/ZnS QDs was achieved, which was 1.7 times higher than that of commercial MRI contrast agent (6.4667 mM–1 S–1). Furthermore, the cellular internalization of Gd:AgInS2/ZnS QDs exerts no significant effect on the adipogenesis of MSCs and is conducive to the observation of further adipogenic differentiation. Our work helps to verify the promising prospect to develop a bimodal nanoprobe of fluorescence/MRI based on Gd:AgInS2/ZnS QDs, which could monitor the differentiation and migration of MSCs for further therapeutic approach.

Development of an Au nanoclusters based activatable nanoprobe for NIR-II fluorescence imaging of gastric acid

Gastric acid is an important functional substance secreted by the stomach of the living organisms, reflecting the gastric physiological condition. The sensing of gastric acid in vivo is of great significance for evaluation of gastric function, diagnosis and treatment of gastric diseases and maintenance of organism health but remains challenging due to the harsh acid and digestive environment of stomach. This study developed an activatable nanoprobe based on Au nanoclusters (Au NCs) for sensitive and real-time noninvasive near-infrared II (NIR-II) fluorescence imaging detection of gastric acid in vivo for the first time. The Au NCs were encapsulated by polydopamine to have enhanced NIR-II luminescence and high stability and combined with methylene blue to possess the pH responsiveness for gastric acid imaging. The developed nanoprobe could not only monitor gastric acid secretion in vivo but also imaging the changes of gastric acid caused by feeding, acid-inhibition drugs and gastric ulcer disease. This study provides a promising avenue for the improvement of the application performance of Au NCs and imaging analysis of gastric acid and related gastric diseases.

Retraction: Design and synthesis of a highly sensitive “Turn-On” fluorescent organic nanoprobe for iron(iii) detection and imaging

Retraction of ‘Design and synthesis of a highly sensitive “Turn-On” fluorescent organic nanoprobe for iron(iii) detection and imaging’ by Cuiping Han et al., J. Mater. Chem. C, 2014, 2, 9077–9082, https://doi.org/10.1039/C4TC01759E.

Engineering molecular nanoprobes to target early atherosclerosis: Precise diagnostic tools and promising therapeutic carriers.

Atherosclerosis, an inflammation-driven chronic blood vessel disease, is a major contributor to devastating cardiovascular events, bringing serious social and economic burdens. Currently, non-invasive diagnostic and therapeutic techniques in combination with novel nanosized materials as well as established molecular targets are under active investigation to develop integrated molecular imaging approaches, precisely visualizing and/or even effectively reversing early-stage plaques. Besides, mechanistic investigation in the past decades provides many potent candidates extensively involved in the initiation and progression of atherosclerosis. Recent hotly-studied imaging nanoprobes for detecting early plaques mainly including optical nanoprobes, photoacoustic nanoprobes, magnetic resonance nanoprobes, positron emission tomography nanoprobes, and other dual- and multi-modality imaging nanoprobes, have been proven to be surface functionalized with important molecular targets, which occupy tailored physical and biological properties for atherogenesis. Of note, these engineering nanoprobes provide long blood-pool residence and specific molecular targeting, which allows efficient recognition of early-stage atherosclerotic plaques and thereby function as a novel type of precise diagnostic tools as well as potential therapeutic carriers of anti-atherosclerosis drugs. There have been no available nanoprobes applied in the clinics so far, although many newly emerged nanoprobes, as exemplified by aggregation-induced emission nanoprobes and TiO2 nanoprobes, have been tested for cell lines in vitro and atherogenic animal models in vivo, achieving good experimental effects. Therefore, there is an urgent call to translate these preclinical results for nanoprobes into clinical trials. For this reason, this review aims to give an overview of currently investigated nanoprobes in the context of atherosclerosis, summarize relevant published studies showing applications of different kinds of formulated nanoprobes in early detection and reverse of plaques, discuss recent advances and some limitations thereof, and provide some insights into the development of the new generation of more precise and efficient molecular nanoprobes, with a critical property of specifically targeting early atherosclerosis.

MRNA-activated DNAzyme nanoprobe for tumor cell precise imaging and gene therapy.

A novel Au-nucleic acid nanoprobe, catalyzed by mRNA, has been developed for live cell imaging and precise treatment of tumor cells. This nanoprobe exhibits the remarkable ability to differentiate between tumor cells and normal cells through live cell mRNA imaging, while selectively inducing apoptosis in tumor cells.

High-fidelity imaging of intracellular microRNA via a bioorthogonal nanoprobe.

The spatiotemporal visualization of intracellular microRNA (miRNA) plays a critical role in the diagnosis and treatment of malignant disease. Although DNAzyme-based biosensing has been regarded as the most promising candidate, inefficient analytical resolution is frequently encountered. Here, we propose a bioorthogonal approach toward high-fidelity imaging of intracellular miRNA by designing a multifunctional nanoprobe that integrates MnO2 nanosheet-mediated intracellular delivery and activation by a fat mass and obesity-associated protein (FTO)-switched positive feedback. MnO2 nanosheets facilitate nanoprobe delivery and intracellular DNAzyme cofactors are released upon glutathione-triggered reduction. Meanwhile, an m6A-caged DNAzyme probe could be bioorthogonally activated by intracellular FTO to eliminate potential off-target activation. Therefore, the activated DNAzyme probe and substrate probe could recognize miRNA to perform cascade signal amplification in the initiation of the release of Mn2+ from MnO2 nanosheets. This strategy realized high-fidelity imaging of intracellular aberrant miRNA within tumor cells with a satisfactory detection limit of 9.7 pM, paving the way to facilitate clinical tumor diagnosis and prognosis monitoring.

A covalent organic framework-derived M1 macrophage mimic nanozyme for precise tumor-targeted imaging and NIR-II photothermal catalytic chemotherapy.

Nanoprobes for efficient tumor-targeted imaging and therapy are urgently needed for clinical tumor theranostics. Herein, inspired by the heterogeneity of the tumor microenvironment, we report a covalent organic framework (COF)-derived biomimetic nanozyme for precise tumor-targeted imaging and NIR-II photothermal-catalysis-enhanced chemotherapy (PTCEC). Using a crystalline nanoscale COF as the precursor, a peroxidase-like porous N-doped carbonous nanozyme (PNC) was obtained, which was cloaked with an M1 macrophage cell membrane (M1m) to create a multifunctional biomimetic nanoprobe for tumor-targeted imaging and therapy. The M1m coating enabled the nanoprobe to target cancer cells and tumor tissues for highly efficient tumor imaging and drug delivery. The peroxidase-like activity of the PNC allowed for the conversion of intratumoral H2O2 into toxic ˙OH that synergized with its NIR-II photothermal effect to strengthen the chemotherapy. Therefore, highly efficient tumor-targeted imaging and NIR-II PTCEC were realized with an M1 macrophage mimic nanoprobe. This work provides a feasible tactic for a biomimetic theranostic nanoprobe and will inspire the development of new bioactive nanomaterials for clinical tumor theranostic applications.

A dye-quenched/sensitized switching upconversion nanoprobe for high-contrast mapping of the pH-related tumor microenvironment.

Nanoprobes based on lanthanide-doped upconversion nanoparticles (UCNPs) exhibit promising potential in bioimaging and biosensing due to their unique optical properties. However, conventional UCNP nanoprobes based on the dye quenching effect are still limited in biosensing due to their low upconversion efficiency. The advent of dye-sensitized upconversion has resulted in nanoprobes with significantly enhanced efficiency; however, these still suffer from a high initial emissive background. In view of this, herein, we have constructed a dye-quenched/sensitized switching upconversion nanoprobe for high-contrast imaging of the pH-related tumor microenvironment. Under normal conditions, the luminescence of the nanoprobe at 540 nm (UCL540) was significantly quenched by the employed dye. Upon being triggered by an acid, the dye would switch to its fluorescent form to sensitize the luminescence of UCNPs, affording a significant enhancement of UCL540. The switching from dye-quenched UCL to dye-sensitized UCL jointly enables the detection of a high signal-to-background ratio (high up to 50), allowing for high-contrast mapping of the tumor specific acidic microenvironment in vivo. We believe that this nanoplatform holds considerable promise for acid-related sensing.

Multifunctional Nanoprobe for 3D Nanoresolution Imaging of Intact Cell HER2 Protein with Hard X-ray Tomography.

Three-dimensional nondestructive, nanoresolution, and in situ visualization of protein spatial localization in a large, thick single cell remains challenging. In this study, we designed a multifunctional iron oxide (Fe@BFK) nanoprobe that possesses fluorescence and hard X-ray imaging signals. This probe can specifically target the human epidermal growth factor receptor 2 (HER2) protein and help optimize the label condition and selection of suitable samples for X-ray imaging. Combining 30 nm resolution synchrotron radiation hard X-ray nanocomputed tomography and the X-ray-sensitive Fe@BFK nanoprobe, a 3D localization of HER2 on SK-BR-3 cells was obtained for the first time. HER2 was mainly localized and cluster-distributed on the cell membrane with a heterogeneous pattern. This study provides a novel method for the in situ and nondestructive synchrotron radiation imaging of the desired protein localization in large, thick cells and evaluation of the true cellular distribution of a nanoprobe with high resolution.

Rational design of a negative photochromic spiropyran-containing fluorescent polymeric nanoprobe for sulfur dioxide derivative ratiometric detection and cell imaging.

It is highly desirable to develop high-performance ratiometric fluorescent probes for SO2 derivative detection and realize their application in biological imaging. In this study, we report the rational design of a novel negative photochromic spiropyran derivative, spiro[azahomoadamantane-pyran] (MAHD-SP), with notable orange fluorescence in its stable ring-opened state without UV regulation. The unsaturated double bond of MAHD-SP underwent the Michael addition reaction of the SO2 derivative, making the fluorescence quenching of MAHD-SP obvious. Then, MAHD-SP, a fluorescent conjugated polymer PFO and a polymeric surfactant PEO113-b-PS49 were used to construct a ratiometric fluorescent polymeric nanoprobe (RFPN) via a coprecipitation method. The probe exhibited high sensitivity and selectivity for the ratiometric detection of SO2 derivatives in pure aqueous solutions. Moreover, the good biocompatibility of RFPN can be used to visualize exogenous and endogenous SO2 derivative generation in living cells.

Predicting thrombolytic haemorrhage risk of acute ischemic stroke through angiogenesis/inflammation dual-targeted MR imaging

Ischemic stroke is currently one of the biggest threats to global health due to its high mortality and disability rate. Thrombolysis with intravenous alteplase remains a primary therapy for acute ischemic stroke. Nevertheless, haemorrhage may be triggered to cause secondary victimization to the patients. On the other hand, the compensatory cerebral collaterals developing in response to ischemia stroke help preserve the penumbral area where the neurons are considered to be salvageable. In this context, precisely evaluating the collateral circulation and predicting the thrombolytic risk of stroke are extremely important not only for treatment decision making but also for a good prognosis. Aiming at identifying the ischemic penumbra and precisely predicting the possible thrombolytic haemorrhage risk prior to thrombolytic therapy, we herein report a dual-targeted magnetic resonance imaging (MRI) nanoprobe based on magnetic iron oxide nanoparticles for visualizing both cerebral collateral circulation and ischemic inflammation. Focal cerebral ischemia rat models were adopted for imaging studies on 7.0 Tesla MRI with different sequences. The imaging results revealed that the nanoprobe could target the newly formed collateral vessels and inflammatory lesions in the cerebral ischemic region, which enabled the visualization of the potential haemorrhage sites of thrombolytic therapy apart from the ischemic penumbra. The current studies therefore offer for the first time a noninvasive approach for precisely predicting potential reperfusion haemorrhage through MRI.