Fluorescence Molecular Tomography System Research Articles

Glucose-Regulated Protein 78 Targeting ICG and DOX Loaded Hollow Fe3O4 Nanoparticles for Hepatocellular Carcinoma Diagnosis and Therapy.

Liver cancer is considered as the third leading cause of cancer-related deaths, with hepatocellular carcinoma (HCC) accounting for approximately 90% of liver cancers. Improving the treatment of HCC is a serious challenge today. The primary objective of this study was to construct SP94-Fe3O4@ICG&DOX nanoparticles and investigate their potential diagnosis and treatment effect benefits on HCC. Firstly, we synthesized and characterized SP94-Fe3O4@ICG&DOX nanoparticles and confirmed their in vitro release behavior, photothermal and photodynamic performance. Moreover, the in vivo imaging capability was also observed. Finally, the inhibitory effects on Hepa1-6 in vitro and in vivo were observed as well as biosafety. SP94-Fe3O4@ICG&DOX nanoparticles have a size of ~22.1 nm, with an encapsulation efficiency of 45.2% for ICG and 42.7% for DOX, showing excellent in vivo MPI and fluorescence imaging capabilities for precise tumor localization, and synergistic photo-chemotherapy (pH- and thermal-sensitive drug release) against tumors under irradiation. With the assistance of a fluorescence molecular imaging system or MPI scanner, the location and contours of the tumor were clearly visible. Under a constant laser irradiation (808 nm, 0.6 W/cm2) and a set concentration (50 µg/mL), the temperature of the solution could rapidly increase to ~45 °C, which could effectively kill the tumor cells. It could be effectively uptaken by HCC cells and significantly inhibit their proliferation under the laser irradiation (100% inhibition rate for HCC tumors). And most importantly, our nanoparticles exhibited favorable biocompatibility with normal tissues and cells. This versatile agent can serve as an intelligent and promising nanoplatform that integrates multiple accurate diagnoses, precise positioning of cancer tissue, and effective coordination with synergistic tumor photodynamic therapy.

Characterization of fluorescence lifetime of organic fluorophores for molecular imaging in the shortwave infrared window

SignificanceFluorescence lifetime imaging in the shortwave infrared (SWIR) is expected to enable high-resolution multiplexed molecular imaging in highly scattering tissue.AimTo characterize the brightness and fluorescence lifetime of commercially available organic SWIR fluorophores and benchmark them against the tail emission of conventional NIR-excited probes.ApproachCharacterization was performed through our established time-domain mesoscopic fluorescence molecular tomography system integrated around a time-correlated single-photon counting-single-photon avalanche diode array. Brightness and fluorescence lifetime were measured for NIR and SWIR probes >1000 nm. Simultaneous probe imaging was then performed to assess their potential for multiplexed studies.ResultsThe NIR probes outperformed SWIR probes in brightness while the mean fluorescence lifetimes of the SWIR probes were extremely short. The phantom study demonstrated the feasibility of lifetime multiplexing in the SWIR window with both NIR and SWIR probes.ConclusionsLong-tail emission of NIR probes outperformed the SWIR probes in brightness >1000 nm. Fluorescence lifetime was readily detectable in the SWIR window, where the SWIR probes showed shorter lifetimes compared to the NIR probes. We demonstrate the feasibility of lifetime multiplexing in the SWIR window, which paves the way for in vivo multiplexed studies of intact tissues at improved resolution.

The Dual-Targeted Peptide Conjugated Probe for Depicting Residual Nasopharyngeal Carcinoma and Guiding Surgery.

Detecting residual nasopharyngeal carcinoma (rNPC) can be difficult because of the coexistence of occult tumours and post-chemoradiation changes, which poses a challenge for both radiologists and surgeons using current imaging methods. Currently, molecular imaging that precisely targets and visualises particular biomarkers in tumours may exceed the specificity and sensitivity of traditional imaging techniques, providing the potential to distinguish tumours from non-neoplastic lesions. Here, we synthesised a HER2/SR-BI-targeted tracer to efficiently position NPC and guide surgery in living mice. This bispecific tracer contained the following two parts: IRDye 800 CW, as an imaging reagent for both optical and optoacoustic imaging, and a fusion peptide (FY-35), as the targeting reagent. Both in vitro and in vivo tests demonstrated that the tracer had higher accumulation and longer retention (up to 48 h) in tumours than a single-targeted probe, and realised sensitive detection of tumours with a minimum size of 3.9 mm. By visualising the vascular network via a customised handheld optoacoustic scan, our intraoperative fluorescence molecular imaging system provides accurate guidance for intraoperative tumour resection. Integrating the advantages of both optical and optoacoustic scanning in an intraoperative image-guided system, this method holds promise for depicting rNPC and guiding salvage surgery.

Non-invasive visualization of amyloid-beta deposits in Alzheimer amyloidosis mice using magnetic resonance imaging and fluorescence molecular tomography.

Abnormal cerebral accumulation of amyloid-beta peptide (Aβ) is a major hallmark of Alzheimer's disease. Non-invasive monitoring of Aβ deposits enables assessing the disease burden in patients and animal models mimicking aspects of the human disease as well as evaluating the efficacy of Aβ-modulating therapies. Previous in vivo assessments of plaque load have been predominantly based on macroscopic fluorescence reflectance imaging (FRI) and confocal or two-photon microscopy using Aβ-specific imaging agents. However, the former method lacks depth resolution, whereas the latter is restricted by the limited field of view preventing a full coverage of the large brain region. Here, we utilized a fluorescence molecular tomography (FMT)-magnetic resonance imaging (MRI) pipeline with the curcumin derivative fluorescent probe CRANAD-2 to achieve full 3D brain coverage for detecting Aβ accumulation in the arcAβ mouse model of cerebral amyloidosis. A homebuilt FMT system was used for data acquisition, whereas a customized software platform enabled the integration of MRI-derived anatomical information as prior information for FMT image reconstruction. The results obtained from the FMT-MRI study were compared to those from conventional planar FRI recorded under similar physiological conditions, yielding comparable time courses of the fluorescence intensity following intravenous injection of CRANAD-2 in a region-of-interest comprising the brain. In conclusion, we have demonstrated the feasibility of visualizing Aβ deposition in 3D using a multimodal FMT-MRI strategy. This hybrid imaging method provides complementary anatomical, physiological and molecular information, thereby enabling the detailed characterization of the disease status in arcAβ mouse models, which can also facilitate monitoring the efficacy of putative treatments targeting Aβ.

Noninvasive multimodal fluorescence and magnetic resonance imaging of whole-organ intervertebral discs.

Low back pain (LBP) is a commonly experienced symptom posing a tremendous healthcare burden to individuals and society at large. The LBP pathology is strongly linked to degeneration of the intervertebral disc (IVD), calling for development of early-stage diagnostic tools for visualizing biomolecular changes in IVD. Multimodal measurements of fluorescence molecular tomography (FMT) and magnetic resonance imaging (MRI) were performed on IVD whole organ culture model using an in-house built FMT system and a high-field MRI scanner. The resulted multimodal images were systematically validated through epifluorescence imaging of the IVD sections at a microscopic level. Multiple image contrasts were exploited, including fluorescence distribution, anatomical map associated with T1-weighted MRI contrast, and water content related with T2 relaxation time. The developed multimodality imaging approach may thus serve as a new assessment tool for early diagnosis of IVD degeneration and longitudinal monitoring of IVD organ culture status using fluorescence markers.

NIR-II/NIR-I Fluorescence Molecular Tomography of Heterogeneous Mice Based on Gaussian Weighted Neighborhood Fused Lasso Method.

Fluorescence molecular tomography (FMT), which can visualize the distribution of fluorescence biomarkers, has become a novel three-dimensional noninvasive imaging technique for in vivo studies such as tumor detection and lymph node location. However, it remains a challenging problem to achieve satisfactory reconstruction performance of conventional FMT in the first near-infrared window (NIR-I, 700-900nm) because of the severe scattering of NIR-I light. In this study, a promising FMT method for heterogeneous mice was proposed to improve the reconstruction accuracy using the second near-infrared window (NIR-II, 1000-1700nm), where the light scattering significantly reduced compared with NIR-I. The optical properties of NIR-II were analyzed to construct the forward model for NIR-II FMT. Furthermore, to raise the accuracy of solution of the inverse problem, we proposed a novel Gaussian weighted neighborhood fused Lasso (GWNFL) method. Numerical simulation was performed to demonstrate the outperformance of GWNFL compared with other algorithms. Besides, a novel NIR-II/NIR-I dual-modality FMT system was developed to contrast the in vivo reconstruction performance between NIR-II FMT and NIR-I FMT. To compare the reconstruction performance of NIR-II FMT with traditional NIR-I FMT, numerical simulations and in vivo experiments were conducted. Both the simulation and in vivo results showed that NIR-II FMT outperformed NIR-I FMT in terms of location accuracy and spatial overlap index. It is believed that this study could promote the development and biomedical application of NIR-II FMT in the future.

Multi-Parametric Standardization of Fluorescence Imaging Systems Based on a Composite Phantom.

Fluorescence molecular imaging (FMI) has emerged as a promising tool for surgical guidance in oncology, with one of the few remaining challenges being the ability to offer quality control and data referencing. This paper investigates the use of a novel composite phantom to correct and benchmark FMI systems. This paper extends on previous work by describing a phantom design that can provide a more complete assessment of FMI systems through quantification of dynamic range and determination of spatial illumination patterns for both reflectance and fluorescence imaging. Various performance metrics are combined into a robust and descriptive "system benchmarking score," enabling not only the comprehensive comparison of different systems, but also for the first time, correction of the acquired data. We show that systems developed for targeted fluorescence imaging can achieve benchmarking scores of up to 70%, while clinically available systems optimized for indocyanine green are limited to 50%, mostly due to greater leakage of ambient and excitation illumination and lower resolution. The image uniformity can also be approximated and employed for image flat-fielding, an important milestone toward data referencing. In addition, we demonstrate composite phantom use in assessing the performance of a surgical microscope and of a raster-scan imaging system. Our results suggest that the new phantom has the potential to support high-fidelity FMI through benchmarking and image correction. Standardization of the FMI is a necessary process for establishing good imaging practices in clinical environments and for enabling high-fidelity imaging across patients and multi-center imaging studies.

Smart Toolkit for Fluorescence Tomography: Simulation, Reconstruction, and Validation.

Fluorescence molecular tomography (FMT) can provide valuable molecular information by mapping the bio-distribution of fluorescent reporter molecules in the intact organism. Various prototype FMT systems have been introduced during the past decade. However, none of them has evolved as a standard tool for routine biomedical research. The goal of this paper is to develop a software package that can automate the complete FMT reconstruction procedure. We present smart toolkit for fluorescence tomography (STIFT), a comprehensive platform comprising three major protocols: 1) virtual FMT, i.e., forward modeling and reconstruction of simulated data; 2) control of actual FMT data acquisition; and 3) reconstruction of experimental FMT data. Both simulation and phantom experiments have shown robust reconstruction results for homogeneous and heterogeneous tissue-mimicking phantoms containing fluorescent inclusions. STIFT can be used for optimization of FMT experiments, in particular for optimizing illumination patterns. This paper facilitates FMT experiments by bridging the gaps between simulation, actual experiments, and data reconstruction.

In vivo long-term investigation of tumor bearing mKate2 by an in-house fluorescence molecular imaging system

BackgroundOptical imaging is one of the most common, low-cost imaging tools used for investigating the tumor biological behavior in vivo. This study explores the feasibility and sensitivity of a near infrared fluorescent protein mKate2 for a long-term non-invasive tumor imaging in BALB/c nude mice, by using a low-power optical imaging system.MethodsIn this study, breast cancer cell line MDA-MB-435s expressing mKate2 and MDA-MB-231 expressing a dual reporter gene firefly luciferase (fLuc)-GFP were used as cell models. Tumor cells were implanted in different animal body compartments including subcutaneous, abdominal and deep tissue area and closely monitored in real-time. A simple and low-power optical imaging system was set up to image both fluorescence and bioluminescence in live animals.ResultsThe presence of malignant tissue was further confirmed by histopathological assay. Considering its lower exposure time and no need of substrate injection, mKate2 is considered a superior choice for subcutaneous imaging compared with fLuc. On the contrary, fLuc has shown to be a better option when monitoring the tumor in a diffusive area such as abdominal cavity. Furthermore, both reporter genes have shown good stability and sensitivity for deep tissue imaging, i.e. tumor within the liver. In addition, fLuc has shown to be an excellent method for detecting tumor cells in the lung.ConclusionsThe combination of mKate2 and fLuc offers a superior choice for long-term non-invasive real-time investigation of tumor biological behavior in vivo.

Image reconstruction in fluorescence molecular tomography with sparsity-initialized maximum-likelihood expectation maximization.

We present a reconstruction method involving maximum-likelihood expectation maximization (MLEM) to model Poisson noise as applied to fluorescence molecular tomography (FMT). MLEM is initialized with the output from a sparse reconstruction-based approach, which performs truncated singular value decomposition-based preconditioning followed by fast iterative shrinkage-thresholding algorithm (FISTA) to enforce sparsity. The motivation for this approach is that sparsity information could be accounted for within the initialization, while MLEM would accurately model Poisson noise in the FMT system. Simulation experiments show the proposed method significantly improves images qualitatively and quantitatively. The method results in over 20 times faster convergence compared to uniformly initialized MLEM and improves robustness to noise compared to pure sparse reconstruction. We also theoretically justify the ability of the proposed approach to reduce noise in the background region compared to pure sparse reconstruction. Overall, these results provide strong evidence to model Poisson noise in FMT reconstruction and for application of the proposed reconstruction framework to FMT imaging.

Optical molecular imaging-guided radiation therapy part 2: Integrated x-ray and fluorescence molecular tomography.

Differentiating tumor from its surrounding soft tissues is challenging for x-ray computed tomography (CT). Fluorescence molecular tomography (FMT) can directly localize the internal tumors targeted with specific fluorescent probes. A FMT system was developed and integrated onto a CT-guided irradiator to improve tumor localization for image-guided radiation. The FMT system was aligned orthogonal to the cone-beam CT onboard our previously developed image-guided small animal arc radiation treatment system (iSMAART). Through rigorous physical registration, the onboard CT provides accurate surface contour which is used to generate three-dimensional mesh for FMT reconstruction. During FMT experiments, a point laser source perpendicular to the rotating axis was used to excite the internal fluorophores. The normalized optical images from multiple projection angles were adopted for tomographic reconstruction. To investigate the accuracy of the FMT in locating the tumor and recovering its volume, in vivo experiments were conducted on two breast cancer models: MDA-MB-231 cancer xenograft on nude mice and 4T1 cancer xenograft on white mice. Both cancer cell lines overexpress the epidermal growth factor receptor (EGFR). A novel fluorescent poly(lactic-co-glycolic) acid (PLGA) nanoparticle conjugated with anti-EGFR was intravenously injected to specifically target the breast cancer cells. Another ex vivo experiment on a mouse bearing a surgically implanted Indocyanine Green-containing glass tube was conducted, to additionally validate the precision of FMT-guided radiation therapy. The FMT can accurately localize the single-nodule breast tumors actively targeted with fluorescent nanoparticles with localization error < 0.5 mm calculated between the centers of mass of tumors in FMT and CT. The reconstructed tumor volume in FMT was significantly correlated with that in the iodinated contrast-enhanced CT (R2 = 0.94, P < 0.001). The FMT was able to guide focal radiation delivery with submillimeter accuracy. Using the tumor-targeting fluorescent probes, the iSMAART with onboard FMT system can accurately differentiate tumors from their surrounding soft tissue, guide precise focal radiation delivery, and potentially assess tumor response in cancer research.

Benchmarking of fluorescence cameras through the use of a composite phantom.

Fluorescence molecular imaging (FMI) has shown potential to detect and delineate cancer during surgery or diagnostic endoscopy. Recent progress on imaging systems has allowed sensitive detection of fluorescent agents even in video rate mode. However, lack of standardization in fluorescence imaging challenges the clinical application of FMI, since the use of different systems may lead to different results from a given study, even when using the same fluorescent agent. In this work, we investigate the use of a composite fluorescence phantom, employed as an FMI standard, to offer a comprehensive method for validation and standardization of the performance of different imaging systems. To exclude user interaction, all phantom features are automatically extracted from the acquired epi-illumination color and fluorescence images, using appropriately constructed templates. These features are then employed to characterize the performance and compare different cameras to each other. The proposed method could serve as a framework toward the calibration and benchmarking of FMI systems, to facilitate their clinical translation.

Method for In-Vivo Fluorescence Imaging Contrast Enhancement through Light Modulation.

Early diagnosis is one of the most important factors that increase the therapeutic potential of the disease. Diagnoses conducted by conventional equipment are expensive, time-consuming, burdensome to patients, and do not have high success rates. Diagnostic methods have also been investigated using nanoparticles. However, there have been no significant improvements in the early diagnosis of disease. The diagnosis technique proposed in this paper consumes less time, is more cost-effective, and more accurate. It uses a new concept-a low-intensity fluorescence molecular imaging system with a lock-in technique. This study applied the lock-in technique to basic research in contrast enhancement and optimization. This improved fluorescence distribution analysis, resulting in increased resolution of optical molecular imaging for early diagnosis of disease. An experimental lock-in fluorescence imaging system, which used a variety of fluorescent dyes, achieved signal amplification 100 times greater than that of a conventional fluorescence imaging system. The results of this study demonstrate that the lock-in technique could significantly improve optical molecular imaging technology, making it possible to achieve early diagnosis of disease.

An Inversion Scheme for Hybrid Fluorescence Molecular Tomography Using a Fuzzy Inference System.

The imaging performance of fluorescence molecular tomography (FMT) improves when information from the underlying anatomy is incorporated into the inversion scheme, in the form of priors. The requirement for incorporation of priors has recently driven the development of hybrid FMT systems coupled to other modalities, such as X-ray CT and MRI. A critical methodological aspect in this modality relates to the particular method selected to incorporate prior information obtained from the anatomical imaging modality into the FMT inversion. We propose herein a new approach for utilizing prior information, which preferentially minimizes residual errors associated with measurements that better describe the anatomical segments considered. This preferential minimization was realized using a weighted least square (WLS) approach, where the weights were optimized using a Mamdani-type fuzzy inference system. The method of priors introduced herein was deployed as a two-step structured regularization approach and was verified with experimental measurements from phantoms as well as ex vivo and in vivo animal studies. The results demonstrate accurate performance and minimization of reconstruction bias, without requiring user input for setting the regularization parameters. As such, the proposed method offers significant progress in incorporation of anatomical priors in FMT and, as a result, in realization of the full potential of hybrid FMT.

Noncontact full-angle fluorescence molecular tomography system based on rotary mirrors.

We propose a novel noncontact fluorescence molecular tomography system that achieves full-angle capacity with the use of a new rotary-mirrors-based imaging head. In the imaging head, four plane mirrors are mounted on a rotating gantry to enable illumination and detection over 360°. In comparison with existing full-angle systems, our system does not require rotation of the specimen animal, a large and heavy light source (with scanning head), or a bulky camera (with filters and lens). The system design and implementation are described in detail. Both physical phantom and in vivo experiments are performed to verify the performance of the proposed system.

A modularly designed fluorescence molecular tomography system for multi-modality imaging.

Nowadays multi-modality imaging has gained great interest in biology research by offering complementary information. In this paper, a modularly designed fluorescence molecular tomography (FMT) system has been developed, which can be not only used as a standalone imaging device, but also feasibly integrated with other imaging modalities, such as X-ray computed tomography (X-CT), single photon emission computed tomography (SPECT) and positron emission tomography (PET), to perform multi-modality imaging in a sequential manner. The system rotates the CCD camera and the excitation light source in the vertical plane, while the animal is stationed on a horizontally moveable transparent animal holder at its natural prone position. FMT and other imaging modalities are co-registered automatically. Phantom and animal experiments have been carried out to demonstrate the performance of the system. The accurate results show that this innovative flexible FMT system has a great potential to be a powerful tool for the study of small animal disease models.

The kinetics of glomerular deposition of nephritogenic IgA.

Whether IgA nephropathy is attributable to mesangial IgA is unclear as there is no correlation between intensity of deposits and extent of glomerular injury and no clear mechanism explaining how these mesangial deposits induce hematuria and subsequent proteinuria. This hinders the development of a specific therapy. Thus, precise events during deposition still remain clinical challenge to clarify. Since no study assessed induction of IgA nephropathy by nephritogenic IgA, we analyzed sequential events involving nephritogenic IgA from IgA nephropathy-prone mice by real-time imaging systems. Immunofluorescence and electron microscopy showed that serum IgA from susceptible mice had strong affinity to mesangial, subepithelial, and subendothelial lesions, with effacement/actin aggregation in podocytes and arcade formation in endothelial cells. The deposits disappeared 24-h after single IgA injection. The data were supported by a fluorescence molecular tomography system and real-time and 3D in vivo imaging. In vivo imaging showed that IgA from the susceptible mice began depositing along the glomerular capillary from 1 min and accumulated until 2-h on the first stick in a focal and segmental manner. The findings indicate that glomerular IgA depositions in IgAN may be expressed under the balance between deposition and clearance. Since nephritogenic IgA showed mesangial as well as focal and segmental deposition along the capillary with acute cellular activation, all glomerular cellular elements are a plausible target for injury such as hematuria.

Limited-projection-angle hybrid fluorescence molecular tomography of multiple molecules.

An advantage of fluorescence methods over other imaging modalities is the ability to concurrently resolve multiple moieties using fluorochromes emitting at different spectral regions. Simultaneous imaging of spectrally separated agents is helpful in interrogating multiple functions or establishing internal controls for accurate measurements. Herein, we investigated multimoiety imaging in the context of a limited-projection-angle hybrid fluorescence molecular tomography (FMT), and x-ray computed tomography implementation and the further registration with positron emission tomography (PET) data. Multichannel FMT systems may image fluorescent probes of varying distribution patterns. Therefore, it is possible that different channels may require different use of priors and regularization parameters. We examined the performance of automatically estimating regularization factors implementing priors, using data-driven regularization specific for limited-projection-angle schemes. We were particularly interested in identifying the implementation variations between hybrid-FMT channels due to probe distribution variation. For this reason, initial validation of the data-driven algorithm on a phantom was followed by imaging different agent distributions in animals, assuming superficial and deep seated activity. We further demonstrate the benefits of combining hybrid FMT with PET to gain multiple readings on the molecular composition of disease.

Research progress in multi-modality imaging system based on fluorescence molecular tomography

Multi-modality imaging system based on fluorescence molecular tomography (FMT) has been widely used in animal studies.It combines FMT with other imaging modalities,which realizes the imaging of anatomical structure,physiological function and biological activities on molecular or cellular levels of small animal in vivo at the same time.This review gives an introduction to the history and current research status of singlemodality FMT system,introduces the developments of multi-modality system based on FMT,especially the system setup,working principle,performance and application of FMT/CT,FMT/MRI and FMT/radionuclide imaging.The prospects of multi-modality imaging system based on FMT are discussed. Key words: Fluorescence molecular tomography; Multi-modality imaging system; FMT/CT; FMT/MRI; FMT/Radionuclide imaging

Dynamic dual-tracer MRI-guided fluorescence tomography to quantify receptor density in vivo

The up-regulation of cell surface receptors has become a central focus in personalized cancer treatment; however, because of the complex nature of contrast agent pharmacokinetics in tumor tissue, methods to quantify receptor binding in vivo remain elusive. Here, we present a dual-tracer optical technique for noninvasive estimation of specific receptor binding in cancer. A multispectral MRI-coupled fluorescence molecular tomography system was used to image the uptake kinetics of two fluorescent tracers injected simultaneously, one tracer targeted to the receptor of interest and the other tracer a nontargeted reference. These dynamic tracer data were then fit to a dual-tracer compartmental model to estimate the density of receptors available for binding in the tissue. Applying this approach to mice with deep-seated gliomas that overexpress the EGF receptor produced an estimate of available receptor density of 2.3 ± 0.5 nM (n = 5), consistent with values estimated in comparative invasive imaging and ex vivo studies.