Multimodal Imaging Probes Research Articles

Facile Preparation of Double Rare Earth-Doped Carbon Dots for MRI/CT/FI Multimodal Imaging

The integration of different imaging functions into an ultrasmall nanoplatform with excellent biocompatibility and metabolic performance to fabricate multimodal imaging probes has been a tremendous challenge. Herein, a novel kind of gadolinium- and ytterbium-doped carbon dots (Gd/Yb@CDs) with diameter of 5.26 ± 0.93 nm was obtained by the convenient one-step hydrothermal process for magnetic resonance imaging (MRI), X-ray computed tomography (CT), and fluorescent imaging (FI). The obtained Gd/Yb@CDs exhibited not only the excitation-dependent emission and superhigh photostability but also higher longitudinal relaxivity (r1 = 6.65 mM–1 s–1) and excellent X-ray absorption performance (45.43 HU L g–1). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, tissue section assessment, and body weight investigation shed light on the superior biocompatibility of these Gd/Yb@CDs. MRI/CT imaging demonstrated these multimodal nanoprobes could give rise to obviously enhanced contrast effect on the x...

Surface Functionalized Multifunctional Gd2O3–Fluorescein Composite Nanorods for Redox Responsive Drug Delivery and Imaging Applications

We have developed multifunctional Gd2O3–fluorescein based magnetic–fluorescent composite nanorods with small, uniform shape and size distribution and excellent colloidal stability for multimodal imaging and redox responsive drug delivery applications. Gd2O3 nanorods have been synthesized via high temperature colloidal route, and composite development has been achieved via one-step polyacrylate coating. This unique coating approach provides controlled surface functional groups along with near zwitterionic surface charge. For redox responsive drug loading, hydrophilic drug daunorubicin has been modified with disulfide linkage and loaded with the composite nanorods via covalent linkage. High drug loading (∼82%) has been observed with excellent redox responsive release behavior. These composite nanorods have been used as multimodal imaging probe, for fluorescence imaging as well as magnetic resonance imaging (MRI). Fluorescence imaging of the composite nanorods inside cancer-positive KB cells exhibit efficien...

Synthesis of Iron Oxide/Gold Composite Nanoparticles Using Polyethyleneimine as a Polymeric Active Stabilizer for Development of a Dual Imaging Probe.

The combination of magnetic and plasmonic properties using iron oxide/gold nanocomposite particles is crucial for the development of multimodal molecular imaging probes. In this study, iron oxide/gold composite nanoparticles (NanoIOGs) were synthesized via the on-site reduction of an Au precursor salt by polyethyleneimine (PEI) molecules attached to iron oxide nanoparticles (IONPs), and they were employed in magnetic resonance and dark-field microscope imaging. PEI is considered as a polymeric active stabilizer (PAS), acting as a reducing agent for the synthesis of Au and a dispersant for nanoparticles. When the IONPs prepared at the PEI concentration of 0.02 wt. % were used for the NanoIOG synthesis, Au nanoseeds were formed around the IONPs. The alloy clusters of IONPs/Au crystals were produced with further reduction depending on PEI concentration. The NanoIOGs exhibited superparamagnetism in a magnetic field and plasmonic response in a dark-field (DF) microscope. The sizes, morphologies, magnetizations, and r2 relaxivities of NanoIOGs were affected significantly by the amount of PEI added during the NanoIOG synthesis. It is suggested that the PAS-mediated synthesis is simple and effective, and can be applied to various nanostructured Au-metal alloys.

GMars-T Enabling Multimodal Subdiffraction Structural and Functional Fluorescence Imaging in Live Cells.

Fluorescent probes with multimodal and multilevel imaging capabilities are highly valuable as imaging with such probes not only can obtain new layers of information but also enable cross-validation of results under different experimental conditions. In recent years, the development of genetically encoded reversibly photoswitchable fluorescent proteins (RSFPs) has greatly promoted the application of various kinds of live-cell nanoscopy approaches, including reversible saturable optical fluorescence transitions (RESOLFT) and stochastic optical fluctuation imaging (SOFI). However, these two classes of live-cell nanoscopy approaches require different optical characteristics of specific RSFPs. In this work, we developed GMars-T, a monomeric bright green RSFP which can satisfy both RESOLFT and photochromic SOFI (pcSOFI) imaging in live cells. We further generated biosensor based on bimolecular fluorescence complementation (BiFC) of GMars-T which offers high specificity and sensitivity in detecting and visualizing various protein-protein interactions (PPIs) in different subcellular compartments under physiological conditions (e.g., 37 °C) in live mammalian cells. Thus, the newly developed GMars-T can serve as both structural imaging probe with multimodal super-resolution imaging capability and functional imaging probe for reporting PPIs with high specificity and sensitivity based on its derived biosensor.

Rational Design of Multifunctional Fe@γ-Fe2 O3 @H-TiO2 Nanocomposites with Enhanced Magnetic and Photoconversion Effects for Wide Applications: From Photocatalysis to Imaging-Guided Photothermal Cancer Therapy.

Titanium dioxide (TiO2 ) has been widely investigated and used in many areas due to its high refractive index and ultraviolet light absorption, but the lack of absorption in the visible-near infrared (Vis-NIR) region limits its application. Herein, multifunctional Fe@γ-Fe2 O3 @H-TiO2 nanocomposites (NCs) with multilayer-structure are synthesized by one-step hydrogen reduction, which show remarkably improved magnetic and photoconversion effects as a promising generalists for photocatalysis, bioimaging, and photothermal therapy (PTT). Hydrogenation is used to turn white TiO2 in to hydrogenated TiO2 (H-TiO2 ), thus improving the absorption in the Vis-NIR region. Based on the excellent solar-driven photocatalytic activities of the H-TiO2 shell, the Fe@γ-Fe2 O3 magnetic core is introduced to make it convenient for separating and recovering the catalytic agents. More importantly, Fe@γ-Fe2 O3 @H-TiO2 NCs show enhanced photothermal conversion efficiency due to more circuit loops for electron transitions between H-TiO2 and γ-Fe2 O3 , and the electronic structures of Fe@γ-Fe2 O3 @H-TiO2 NCs are calculated using the Vienna ab initio simulation package based on the density functional theory to account for the results. The reported core-shell NCs can serve as an NIR-responsive photothermal agent for magnetic-targeted photothermal therapy and as a multimodal imaging probe for cancer including infrared photothermal imaging, magnetic resonance imaging, and photoacoustic imaging.

Emerging applications of near-infrared fluorescent metal nanoclusters for biological imaging

Fluorescent metal nanoclusters (MNCs) have recently emerged as a novel kind of promising fluorescent probes for biological imaging because of their ultrasmall core size (<2nm), strong photoluminescence, facile availability and good biocompatibility. In this review, we provide an update on recent advances in the development of near infrared (NIR)-emitting MNCs in terms of synthesis strategies and bioimaging applications. We mainly focus on the utilization of NIR-emitting MNCs (including Au, Ag, Cu and alloy NCs) either as single modal imaging (fluorescence intensity-based imaging, fluorescence lifetime imaging, two-photon imaging) probes or as multimodal imaging (such as NIR fluorescence/X-ray computed tomography/magnetic resonance imaging, NIR fluorescence/photoacoustic imaging/magnetic resonance imaging, NIR fluorescence/single photon emission computed tomography) probes in biological cells and tissues. Finally, we give a brief outlook on the future challenges and prospects of developing NIR-emitting MNCs for bioimaging.

PET-MR and SPECT-MR multimodality probes: Development and challenges.

Positron emission tomography (PET)-magnetic resonance (MR) or single photon emission computed tomography (SPECT)-MR hybrid imaging is being used in daily clinical practice. Due to its advantages over stand-alone PET, SPECT or MR imaging, in many areas such as oncology, the demand for hybrid imaging techniques is increasing dramatically. The use of multimodal imaging probes or biomarkers in a single molecule or particle to characterize the imaging subjects such as disease tissues certainly provides us with more accurate diagnosis and promotes therapeutic accuracy. A limited number of multimodal imaging probes are being used in preclinical and potential clinical investigations. The further development of multimodal PET-MR and SPECT-MR imaging probes includes several key elements: novel synthetic strategies, high sensitivity for accurate quantification and high anatomic resolution, favourable pharmacokinetic profile and target-specific binding of a new probe. This review thoroughly summarizes all recently available and noteworthy PET-MR and SPECT-MR multimodal imaging probes including small molecule bimodal probes, nano-sized bimodal probes, small molecular trimodal probes and nano-sized trimodal probes. To the best of our knowledge, this is the first comprehensive overview of all PET-MR and SPECT-MR multimodal probes. Since the development of multimodal PET-MR and SPECT-MR imaging probes is an emerging research field, a selection of 139 papers were recognized following the literature review. The challenges for designing multimodal probes have also been addressed in order to offer some future research directions for this novel interdisciplinary research field.

An all-in-one nanoparticle (AION) contrast agent for breast cancer screening with DEM-CT-MRI-NIRF imaging.

Conventional X-ray mammography has low diagnostic sensitivity for women with dense breasts. As a result, alternative contrast-enhanced screening tools such as dual energy mammography (DEM), computed tomography (CT), magnetic resonance imaging (MRI), and near-infrared fluorescence (NIRF) imaging are being used or investigated for these women. However, currently available contrast agents are non-ideal, have safety issues, and each imaging technique requires a different contrast agent. We therefore sought to develop a multimodal contrast agent that is functional for each breast imaging modality to simplify the diagnosis process and address the issues of existing contrast agents. Herein, we present a novel "all-in-one" nanoparticle (AION) multimodal imaging probe that has potent DEM, CT, MRI, and NIRF contrast properties and improved biocompatibility. AION were formed by co-encapsulation of a near-infrared fluorophore (DiR), silver sulfide nanoparticles (Ag2S-NP), and iron oxide nanoparticles (IO-NP) in PEGylated micelles. AION showed negligible cytotoxicity, which was in agreement with its minimal silver ion release profiles. AION generated strong contrast with all imaging modalities as demonstrated in phantom imaging. AION allowed in vivo tumor imaging as evidenced by the increase in contrast after injection. This study indicates the potential of AION as an effective multimodal contrast agent for breast cancer diagnosis with a range of imaging methods.

Firefly Luciferin-Inspired Biocompatible Chemistry for Protein Labeling and In Vivo Imaging.

Biocompatible reactions have emerged as versatile tools to build various molecular imaging probes that hold great promise for the detection of biological processes in vitro and/or in vivo. In this Minireview, we describe the recent advances in the development of a firefly luciferin-inspired biocompatible reaction between cyanobenzothiazole (CBT) and cysteine (Cys), and highlight its versatility to label proteins and build multimodality molecular imaging probes. The review starts from the general introduction of biocompatible reactions, which is followed by briefly describing the development of the firefly luciferin-inspired biocompatible chemistry. We then discuss its applications for the specific protein labeling and for the development of multimodality imaging probes (fluorescence, bioluminescence, MRI, PET, photoacoustic, etc.) that enable high sensitivity and spatial resolution imaging of redox environment, furin and caspase-3/7 activity in living cells and mice. Finally, we offer the conclusions and our perspective on the various and potential applications of this reaction. We hope that this review will contribute to the research of biocompatible reactions for their versatile applications in protein labeling and molecular imaging.

Chelator-Free Labeling of Metal Oxide Nanostructures with Zirconium-89 for Positron Emission Tomography Imaging.

Radiolabeling of molecules or nanoparticles to form imaging probes is critical for positron emission tomography (PET) imaging, which, with high sensitivity and the ability for quantitative imaging, has been widely used in the clinic. While conventional radiolabeling often employs chelator molecules, a general method for chelator-free radiolabeling of a wide range of materials remains to be developed. Herein, we determined that 10 different types of metal oxide (MxOy, M = Gd, Ti, Te, Eu, Ta, Er, Y, Yb, Ce, or Mo, x = 1-2, y = 2-5) nanomaterials with polyethylene glycol (PEG) modification could be labeled with 89Zr, a PET tracer, via a simple yet general chelator-free radiolabeling method upon simple mixing. High-labeling yields and good serum stabilities are achieved with this method, owing to the strong bonding between oxyphilic 89Zr4+ with oxygen atoms on the MxOy surface. Selecting 89Zr-Gd2O3-PEG as a multimodal imaging probe, we have successfully demonstrated in vivo PET imaging of draining lymph nodes, which are also visualized under magnetic resonance imaging, showing advantages over free 89Zr in the mapping of draining lymph node networks. Our work describes a general and simple method for chelator-free radiolabeling of metal oxide nanostructures, which is promising for the development of multifunctional nanoprobes in biomedical imaging.

Advanced Methods for Radiolabeling Multimodality Nanomedicines for SPECT/MRI and PET/MRI.

The advent of hybrid cameras that combine MRI with either SPECT or PET has stimulated growing interest in developing multimodality imaging probes. Countless options are available for fusing magnetically active species with positron- or γ-ray-emitting radionuclides. The initial problem is one of choice: which chemical systems are a suitable basis for developing hybrid imaging agents? Any attempt to answer this question must also address how the physical, chemical, and biologic properties of a unified imaging agent can be tailored to ensure that optimum specificity and contrast are achieved simultaneously for both imaging modalities. Nanoparticles have emerged as attractive platforms for building multimodality radiotracers for SPECT/MRI and PET/MRI. A wide variety of nanoparticle constructs have been utilized as radiotracers, but irrespective of the particle class, radiolabeling remains a key step. Classic methods for radiolabeling nanoparticles involve functionalization of the particle surface, core, or coating. These modifications typically rely on using traditional metal ion chelate or prosthetic group chemistries. Though seemingly innocuous, appending nanoparticles with these radiolabeling handles can have dramatic effects on important properties such as particle size, charge, and solubility. In turn, alterations in the chemical and physical properties of the nanoparticle often have a negative impact on their pharmacologic profile. A central challenge in radiolabeling nanoparticles is to identify alternative chemical methods that facilitate the introduction of a radioactive nuclide without detrimental effects on the pharmacokinetic and toxicologic properties of the construct. Efforts to solve this challenge have generated a range of innovative chelate-free radiolabeling methods that exploit intrinsic chemical features of nanoparticles. Here, the chemistry of 9 mechanistically distinct methods for radiolabeling nanoparticles is presented. This discourse illustrates the evolution of nanoparticle radiochemistry from classic approaches to modern chelate-free or intrinsic methods.

Molecular Imaging Using Supramolecular Nanotube Constructs

In the September issue of Chem , Pascu and co-workers report their research on the application of supramolecular constructs to the formation of multimodality imaging probes. Basing their work on single-walled carbon nanotubes, they encapsulated radioactive metal ions, sealed the ends of the tubes with fullerenes, and coated them with polysaccharides. The resulting construct could be imaged on a cellular level by multiphoton fluorescence imaging and on a whole animal basis by positron emission tomography.

Facile synthesis of Gd-doped CdTe quantum dots with optimized properties for optical/MR multimodal imaging.

Each imaging modality has its own merits and intrinsic limitations; therefore, combining two or more complementary imaging modalities has become an interesting area of research. Recently, magnetic ion-doped quantum dots have become an increasingly promising class of optical/magnetic resonance multimodal imaging probes due to their excellent physical and chemical properties. In this work, Gd-doped CdTe quantum dots (QDs) were successfully synthesized via a facile one-step refluxing route,and their optimal synthesis conditions were investigated. The prepared CdTe:Gd QDs were shown to exhibit good optical properties with high quantum yields up to 69%, high longitudinal relaxivity (r 1=3.8mM-1s-1), and good crystalline structures. In addition, after further QD surface modification with dextran amine (DA), the resulting DA-modified QDs (i.e. DA-CdTe:Gd QDs) showed strong magnetic resonance imaging contrast (r 1=3.5mM-1s-1) and improved biocompatibility when tested with cell cultures in vitro. Taken together, this new material demonstrated promising performances for both optical and magnetic resonance imaging modalities, suggesting its promising potential applications in non-invasive imaging, particularly in neuronal tracing.

Single ultrasmall Mn2+-doped NaNdF4 nanocrystals as multimodal nanoprobes for magnetic resonance and second near-infrared fluorescence imaging

Multimodal imaging probes have attracted wide attention and have potential to diagnose diseases accurately because of the complementary advantages of multiple imaging modalities. However, intractable issues remain with regard to their complicated multi-step fabrication for hybrid nanostructure and interference of different modal imaging. In the present study, we present, for the first time, T1 and T2-weighted magnetic resonance imaging (MRI) of ultrasmall Mn2+-doped NaNdF4 nanocrystals (NCs), which can also be used simultaneously for second near infrared (NIR-II) fluorescence and computed tomography (CT) imaging, thus enabling high-performance multimodal MRI/NIR-II/CT imaging of single NaNdF4:Mn NCs. The NaNdF4:Mn was demonstrated as a nanoprobe for in vitro and in vivo multimodal MRI and NIR-II fluorescence imaging of human mesenchymal stem cells. The results provide a new strategy to simplify the nanostructure and preparation of probes, based on the features of NaNdF4:Mn NCs, which offer highly efficient multimodal MRI/NIR-II/CT imaging.

Radioiodination of BODIPY and its application to a nuclear and optical dual functional labeling agent for proteins and peptides

In molecular imaging research, the development of multimodal imaging probes has recently attracted much attention. In the present study, we prepared radioiodinated BODIPY and applied it as a nuclear and optical dual functional labeling agent for proteins and peptides. We designed and synthesized [125I]BODIPY with a N-hydroxysuccinimide (NHS) ester, and evaluated its utility as a nuclear and fluorescent dual labeling agent for proteins and peptides. In the radioiodination reaction of BODIPY-NHS with [125I]NaI, [125I]BODIPY-NHS was obtained at a 48% radiochemical yield. When we carried out the conjugation reaction of [125I]BODIPY-NHS with bovine serum albumin (BSA) and RGD (Arg-Gly-Asp) peptide as a model protein and peptide, respectively, [125I]BODIPY-BSA and [125I]BODIPY-RGD peptide were successfully prepared at 98 and 82% radiochemical yields, respectively. Furthermore, we prepared [123I]BODIPY-trastuzumab by this conjugation reaction and successfully applied it to single photon emission computed tomography (SPECT) imaging studies using tumor-bearing mice, suggesting that radioiodinated BODIPY-NHS serves as a dual functional labeling agent for proteins and peptides. Since iodine has various radioisotopes that can be used for SPECT and positron emission tomography (PET) imaging, biological research, and radiotherapy, the radioiodinated BODIPY may be extensively applicable from basic to clinical research.

NIR-to-NIR UCL/T1-weighted MR/CT multimodal imaging by NaYbF4:Tm@NaGdF4:Yb-PVP upconversion nanoparticles

Multimodal imaging nanoprobes are urgently sought because they can integrate different imaging function into individual nanoplatform and provide more comprehensive and accurate information for the diagnosis of early-stage tumor. Lanthanide-based upconversion nanoparticles (UCNPs) are regarded as promising nanoplatforms to fabricate these probes. Herein, we firstly developed the active core-active shell structured NaYbF4:Tm@NaGdF4:Yb-PVP UCNPs with the average diameter of 13.23±0.96nm as multimodal imaging probes. These water-dispersible nanoprobes presented excellent near-infrared to near-infrared (NIR-to-NIR) upconversion luminescence (UCL) performance, which is favorable for optical bioimaging due to deeper tissue penetration and autofluorescence reduction. After coated with the NaGdF4:Yb active shell, the UCL emission intensity at 800nm increased by 7.2times. These nanoprobes exhibited a desirable longitudinal relaxivity (r1=3.58L/(mmols)) and strong X-ray attenuation property (58.84HUL/g). The cytotoxicity assessment, histology analysis and biodistribution study revealed that NaYbF4:Tm@NaGdF4:Yb-PVP UCNPs had relatively low cytotoxicity and negligible organ toxicity. These UCNPs were applied for NIR-to-NIR UCL imaging in vivo. More importantly, the detection of small tumor was successfully achieved under T1-weighted MRI and CT imaging modalities after intravenous injection of these UCNPs. These results revealed that NaYbF4:Tm@NaGdF4:Yb-PVP UCNPs could serve as promising NIR-to-NIR UCL/MRI/CT trimodal imaging probes.

Functional magnetic hybrid nanomaterials for biomedical diagnosis and treatment.

Magnetic nanomaterials integrating supplemental functional materials are called magnetic hybrid nanomaterials (MHNs). Such MHNs have drawn increasing attention due to their biocompatibility and the potential applications either as alternative contrast enhancing agents or effective heat nanomediators in hyperthermia therapy. The joint function comes from the hybrid nanostructures. Hybrid nanostructures of different modification can be easily achieved owing to the large surface-area-to-volume ratio and sophisticated surface characteristic. In this focus article, we mainly discussed the design and synthesis of MHNs and their applications as multimodal imaging probes and therapy agents in biomedicine. These MHNs consisting magnetic nanomaterials with functional nanocomponents such as noble metal or isotopes could perform not only superparamagnetism but also features that can be adapted in, for example, enhancing computed tomography contrast modalities, positron emission tomography, and single-photon emission computed tomography. The combination of several techniques provides more comprehensive information by both synergizing the advantages, such as quantitative evaluation, higher sensitivity and spatial resolution, and mitigating the disadvantages. Such hybrid nanostructures could also provide a unique nanoplatform for enhanced medical tracing, magnetic field, and light-triggered hyperthermia. Moreover, potential advantages and opportunities will be achieved via a combination of diagnostic and therapeutic agents within a single platform, which is so-called 'theranostics.' We expect the combination of unique structural characteristics and integrated functions of multicomponent magnetic hybrid nanomaterials will attract increasing research interest and could lead to new opportunities in nanomedicine and nanobiotechnology. WIREs Nanomed Nanobiotechnol 2018, 10:e1476. doi: 10.1002/wnan.1476 This article is categorized under: Diagnostic Tools > Diagnostic Nanodevices.

Hybrid Core-Shell (HyCoS) Nanoparticles produced by Complex Coacervation for Multimodal Applications

Multimodal imaging probes can provide diagnostic information combining different imaging modalities. Nanoparticles (NPs) can contain two or more imaging tracers that allow several diagnostic techniques to be used simultaneously. In this work, a complex coacervation process to produce core-shell completely biocompatible polymeric nanoparticles (HyCoS) for multimodal imaging applications is described. Innovations on the traditional coacervation process are found in the control of the reaction temperature, allowing a speeding up of the reaction itself, and the production of a double-crosslinked system to improve the stability of the nanostructures in the presence of a clinically relevant contrast agent for MRI (Gd-DTPA). Through the control of the crosslinking behavior, an increase up to 6 times of the relaxometric properties of the Gd-DTPA is achieved. Furthermore, HyCoS can be loaded with a high amount of dye such as ATTO 633 or conjugated with a model dye such as FITC for in vivo optical imaging. The results show stable core-shell polymeric nanoparticles that can be used both for MRI and for optical applications allowing detection free from harmful radiation. Additionally, preliminary results about the possibility to trigger the release of a drug through a pH effect are reported.

A Dual-Modal SERS/Fluorescence Gold Nanoparticle Probe for Mitochondrial Imaging.

A novel SERS/fluorescent multimodal imaging probe for mitochondria has been synthesised using 12 nm diameter gold nanoparticles (AuNP) surface functionalised with a rhodamine thiol derivative ligand. The normal pH-dependent fluorescence of the rhodamine-based ligand is inversed when it is conjugated with the AuNP and higher emission intensity is observed at basic pH. This switch correlates to a pKa at pH 6.62, which makes it an ideal candidate for a pH-sensitive imaging probe in the biological range (pH 6.5-7.4). The observed pH sensitivity of the ligand when attached to the AuNP is thought to be due to the formation of a spirolactam ring, going from positively charged (+18 mV) to negatively charged (-60 mV) as the pH is changed from acidic to basic. Additionally, conjugation of the ligand to the AuNP serves to enhance the Raman signal of the rhodamine ligand through surface-enhanced Raman scattering (SERS). Confocal microscopy has shown that the probe enters HEK293 (kidney), A2780 (ovarian cancer) and Min6 (pancreatic beta) cells within an hour and a half incubation time. The probe was shown to localise in the mitochondria, thus providing a novel pH-dependent SERS/fluorescent multimodal imaging probe for mitochondria.

18F-positron-emitting/fluorescent labeled erythrocytes allow imaging of internal hemorrhage in a murine intracranial hemorrhage model

An agent for visualizing cells by positron emission tomography is described and used to label red blood cells. The labeled red blood cells are injected systemically so that intracranial hemorrhage can be visualized by positron emission tomography (PET). Red blood cells are labeled with 0.3 µg of a positron-emitting, fluorescent multimodal imaging probe, and used to non-invasively image cryolesion induced intracranial hemorrhage in a murine model (BALB/c, 2.36 × 108 cells, 100 µCi, <4 mm hemorrhage). Intracranial hemorrhage is confirmed by histology, fluorescence, bright-field, and PET ex vivo imaging. The low required activity, minimal mass, and high resolution of this technique make this strategy an attractive alternative for imaging intracranial hemorrhage. PET is one solution to a spectrum of issues that complicate single photon emission computed tomography (SPECT). For this reason, this application serves as a PET alternative to [99mTc]-agents, and SPECT technology that is used in 2 million annual medical procedures. PET contrast is also superior to gadolinium and iodide contrast angiography for its lack of clinical contraindications.