Thomas C. Basse-Lüsebrink

Visualization of abscess formation in a murine thigh infection model of Staphylococcus aureus by 19F-magnetic resonance imaging (MRI).

Background During the last years, 19F-MRI and perfluorocarbon nanoemulsion (PFC) emerged as a powerful contrast agent based MRI methodology to track cells and to visualize inflammation. We applied this new modality to visualize deep tissue abscesses during acute and chronic phase of inflammation caused by Staphylococcus aureus infection. Methodology and Principal Findings In this study, a murine thigh infection model was used to induce abscess formation and PFC or CLIO (cross linked ironoxides) was administered during acute or chronic phase of inflammation. 24 h after inoculation, the contrast agent accumulation was imaged at the site of infection by MRI. Measurements revealed a strong accumulation of PFC at the abscess rim at acute and chronic phase of infection. The pattern was similar to CLIO accumulation at chronic phase and formed a hollow sphere around the edema area. Histology revealed strong influx of neutrophils at the site of infection and to a smaller extend macrophages during acute phase and strong influx of macrophages at chronic phase of inflammation. Conclusion and Significance We introduce 19F-MRI in combination with PFC nanoemulsions as a new platform to visualize abscess formation in a murine thigh infection model of S. aureus. The possibility to track immune cells in vivo by this modality offers new opportunities to investigate host immune response, the efficacy of antibacterial therapies and the influence of virulence factors for pathogenesis.

Visualization of inflammation using 19F‐magnetic resonance imaging and perfluorocarbons

Inflammation plays a central pathophysiological role in a large number of diseases. While conventional magnetic resonance imaging (MRI) can depict gross tissue alterations due to proton changes, specific visualization of inflammation is an unmet task in clinical medicine. (19) F/(1) H MRI is a novel technology that allows tracking of stem and immune cells in experimental disease models after labelling with perfluorocarbon (PFC) emulsions. (19) F markers such as PFC compounds provide a unique signal in vivo due to the negligible (19) F background signal of the body. Concomitant acquisition of (1) H images places the labelled cells into their anatomical context. This novel imaging technique has been applied to monitor immune cell responses in myocardial infarction, pneumonia, bacterial abscess formation, peripheral nerve injury, and rejection of donor organs after transplantation. Upon systemic application PFC nanoparticles are preferentially phagozytosed by circulating monocytes/macrophages and, thus, the fluorine signal in inflamed organs mainly reflects macrophage infiltration. Moreover, attenuation of the inflammatory response after immunosuppressive or antibiotic treatments could be depicted based on (19) F/(1) H-MRI. Compared to other organ systems (19) F-MRI of neuroinflammation is still challenging, mainly because of lack in sensitivity. In focal cerebral ischemia early application of PFCs revealed ongoing thrombotic vessel occlusion rather than cell migration indicating that timing of contrast agent application is critical. Current restrictions of (19) F/(1) H-MRI in sensitivity may be overcome by improved imaging hardware, imaging sequences and reconstruction techniques, as well as improved label development and cell labelling procedures in the future.

Application of compressed sensing to in vivo 3D 19F CSI

This study shows how applying compressed sensing (CS) to (19)F chemical shift imaging (CSI) makes highly accurate and reproducible reconstructions from undersampled datasets possible. The missing background signal in (19)F CSI provides the required sparsity needed for application of CS. Simulations were performed to test the influence of different CS-related parameters on reconstruction quality. To test the proposed method on a realistic signal distribution, the simulation results were validated by ex vivo experiments. Additionally, undersampled in vivo 3D CSI mouse datasets were successfully reconstructed using CS. The study results suggest that CS can be used to accurately and reproducibly reconstruct undersampled (19)F spectroscopic datasets. Thus, the scanning time of in vivo(19)F CSI experiments can be significantly reduced while preserving the ability to distinguish between different (19)F markers. The gain in scan time provides high flexibility in adjusting measurement parameters. These features make this technique a useful tool for multiple biological and medical applications.

Monitoring of Monocyte Recruitment in Reperfused Myocardial Infarction With Intramyocardial Hemorrhage and Microvascular Obstruction by Combined Fluorine 19 and Proton Cardiac Magnetic Resonance Imaging

Background— Monocytes and macrophages are indispensable in the healing process after myocardial infarction (MI); however, the spatiotemporal distribution of monocyte infiltration and its correlation to prognostic indicators of reperfused MI have not been well described. Methods and Results— With combined fluorine 19/proton (1H) magnetic resonance imaging, we noninvasively visualized the spatiotemporal recruitment of monocytes in vivo in a rat model of reperfused MI. Blood monocytes were labeled by intravenous injection of 19F-perfluorocarbon emulsion 1 day after MI. The distribution patterns of monocyte infiltration were correlated to the presence of microvascular obstruction (MVO) and intramyocardial hemorrhage. In vivo, 19F/1H magnetic resonance imaging performed in series revealed that monocyte infiltration was spatially inhomogeneous in reperfused MI areas. In the absence of MVO, monocyte infiltration was more intense in MI regions with serious ischemia-reperfusion injuries, indicated by severe intramyocardial hemorrhage; however, monocyte recruitment was significantly impaired in MVO areas accompanied by severe intramyocardial hemorrhage. Compared with MI with isolated intramyocardial hemorrhage, MI with MVO resulted in significantly worse pump function of the left ventricle 28 days after MI. Conclusions— Monocyte recruitment was inhomogeneous in reperfused MI tissue. It was highly reduced in MVO areas defined by magnetic resonance imaging. The impaired monocyte infiltration in MVO regions could be related to delayed healing and worse functional outcomes in the long term. Therefore, monocyte recruitment in MI with MVO could be a potential diagnostic and therapeutic target that could be monitored noninvasively and longitudinally by 19F/1H magnetic resonance imaging in vivo.

Monitoring of Monocyte Recruitment in Reperfused Myocardial Infarction with Intramyocardial Hemorrhage and Microvascular Obstruction by Combined Fluorine-19 and Proton Cardiac MRI

Background— Monocytes and macrophages are indispensable in the healing process after myocardial infarction (MI); however, the spatiotemporal distribution of monocyte infiltration and its correlation to prognostic indicators of reperfused MI have not been well described. Methods and Results— With combined fluorine 19/proton (1H) magnetic resonance imaging, we noninvasively visualized the spatiotemporal recruitment of monocytes in vivo in a rat model of reperfused MI. Blood monocytes were labeled by intravenous injection of 19F-perfluorocarbon emulsion 1 day after MI. The distribution patterns of monocyte infiltration were correlated to the presence of microvascular obstruction (MVO) and intramyocardial hemorrhage. In vivo, 19F/1H magnetic resonance imaging performed in series revealed that monocyte infiltration was spatially inhomogeneous in reperfused MI areas. In the absence of MVO, monocyte infiltration was more intense in MI regions with serious ischemia-reperfusion injuries, indicated by severe intramyocardial hemorrhage; however, monocyte recruitment was significantly impaired in MVO areas accompanied by severe intramyocardial hemorrhage. Compared with MI with isolated intramyocardial hemorrhage, MI with MVO resulted in significantly worse pump function of the left ventricle 28 days after MI. Conclusions— Monocyte recruitment was inhomogeneous in reperfused MI tissue. It was highly reduced in MVO areas defined by magnetic resonance imaging. The impaired monocyte infiltration in MVO regions could be related to delayed healing and worse functional outcomes in the long term. Therefore, monocyte recruitment in MI with MVO could be a potential diagnostic and therapeutic target that could be monitored noninvasively and longitudinally by 19F/1H magnetic resonance imaging in vivo.

Magnetic Resonance Imaging of Tumors Colonized with Bacterial Ferritin-Expressing Escherichia coli

Background Recent studies have shown that human ferritin can be used as a reporter of gene expression for magnetic resonance imaging (MRI). Bacteria also encode three classes of ferritin-type molecules with iron accumulation properties. Methods and Findings Here, we investigated whether these bacterial ferritins can also be used as MRI reporter genes and which of the bacterial ferritins is the most suitable reporter. Bacterial ferritins were overexpressed in probiotic E. coli Nissle 1917. Cultures of these bacteria were analyzed and those generating highest MRI contrast were further investigated in tumor bearing mice. Among members of three classes of bacterial ferritin tested, bacterioferritin showed the most promise as a reporter gene. Although all three proteins accumulated similar amounts of iron when overexpressed individually, bacterioferritin showed the highest contrast change. By site-directed mutagenesis we also show that the heme iron, a unique part of the bacterioferritin molecule, is not critical for MRI contrast change. Tumor-specific induction of bacterioferritin-expression in colonized tumors resulted in contrast changes within the bacteria-colonized tumors. Conclusions Our data suggest that colonization and gene expression by live vectors expressing bacterioferritin can be monitored by MRI due to contrast changes.

In Vivo Imaging of Stepwise Vessel Occlusion in Cerebral Photothrombosis of Mice by 19F MRI

Background 19F magnetic resonance imaging (MRI) was recently introduced as a promising technique for in vivo cell tracking. In the present study we compared 19F MRI with iron-enhanced MRI in mice with photothrombosis (PT) at 7 Tesla. PT represents a model of focal cerebral ischemia exhibiting acute vessel occlusion and delayed neuroinflammation. Methods/Principal Findings Perfluorocarbons (PFC) or superparamagnetic iron oxide particles (SPIO) were injected intravenously at different time points after photothrombotic infarction. While administration of PFC directly after PT induction led to a strong 19F signal throughout the entire lesion, two hours delayed application resulted in a rim-like 19F signal at the outer edge of the lesion. These findings closely resembled the distribution of signal loss on T2-weighted MRI seen after SPIO injection reflecting intravascular accumulation of iron particles trapped in vessel thrombi as confirmed histologically. By sequential administration of two chemically shifted PFC compounds 0 and 2 hours after illumination the different spatial distribution of the 19F markers (infarct core/rim) could be visualized in the same animal. When PFC were applied at day 6 the fluorine marker was only detected after long acquisition times ex vivo. SPIO-enhanced MRI showed slight signal loss in vivo which was much more prominent ex vivo indicative for neuroinflammation at this late lesion stage. Conclusion Our study shows that vessel occlusion can be followed in vivo by 19F and SPIO-enhanced high-field MRI while in vivo imaging of neuroinflammation remains challenging. The timing of contrast agent application was the major determinant of the underlying processes depicted by both imaging techniques. Importantly, sequential application of different PFC compounds allowed depiction of ongoing vessel occlusion from the core to the margin of the ischemic lesions in a single MRI measurement.

Fast CPMG‐based Bloch‐Siegert B1+ mapping

A novel method for B  1+ mapping based on the Bloch‐Siegert (BS) shift was recently presented. This method applies off‐resonant pulses before signal acquisition to encode B1 information into the signal phase. BS‐based methods possess significant advantages in measurement time and accuracy compared to magnitude‐based B  1+ methods. This study extends the idea of BS B  1+ mapping to Carr, Purcell, Meiboom, Gill (CPMG)‐based multi‐spin‐echo (BS‐CPMG‐MSE) and turbo‐spin‐echo (BS‐CPMG‐TSE) imaging. Compared to BS‐based spin echo imaging (BS‐SE), faster acquisition of the B  1+ information was possible using the BS‐CPMG‐TSE sequence. Furthermore, signal loss by T2* effects could be minimized using these spin echo‐based techniques. These effects are critical for gradient echo‐based BS methods at high field strengths. However, multi‐spin‐echo‐based BS B1 methods inherently possess high specific absorption rates. Thus, the relative specific absorption rate of BS‐CPMG‐TSE sequences was estimated and compared with the specific absorption rate produced by BS‐SE sequences. Magn Reson Med, 2012.

Compressed sensing for reduction of noise and artefacts in direct PET image reconstruction

AIM Image reconstruction in positron emission tomography (PET) can be performed using either direct or iterative methods. Direct reconstruction methods need a short reconstruction time. However, for data containing few counts, they often result in poor visual images with high noise and reconstruction artefacts. Iterative reconstruction methods such as ordered subset expectation maximization (OSEM) can lead to overestimation of activity in cold regions distorting quantitative analysis. The present work investigates the possibilities to reduce noise and reconstruction artefacts of direct reconstruction methods using compressed sensing (CS). MATERIALS AND METHODS Raw data are generated either using Monte Carlo simulations using GATE or are taken from PET measurements with a Siemens Inveon small-animal PET scanner. The fully sampled dataset was reconstructed using filtered backprojection (FBP) and reduced in Fourier space by multiplication with an incoherently undersampled sampling pattern, followed by an additional reconstruction with CS. Different sampling patterns are used and an average of the reconstructions is taken. The images are compared to the results of an OSEM reconstruction and quantified using signal-to-noise ratio (SNR). RESULTS The application of the proposed CS post-processing technique clearly improves the image contrast. Dependent on the undersampling factor, noise and artefacts are reduced resulting in an SNR that is increased up to 3.4-fold. For short acquisition times with low count statistics the SNR of the CS reconstructed image exceeds the SNR of the OSEM reconstruction. CONCLUSION Especially for low count data, the proposed CS-based post-processing method applied to FBP reconstructed PET images enhances the image quality significantly.

SAR-reduced spin-echo-based Bloch–Siegert B1+ mapping: BS-SE-BURST

Fast and accurate B1+ mapping is possible using phase‐based Bloch–Siegert (BS) methods. Importantly, the off‐resonant pulses needed for BS B1+ mapping methods can easily be implemented in multiple MR sequences. BS‐based B1+ mapping has thus been introduced for gradient echo (BS‐FLASH), spin‐echo (BS‐SE), and Carr, Purcell, Meiboom, Gill (CPMG)‐based multi‐SE and turbo‐SE sequences. When using SE and multi‐SE/turbo‐SE‐based BS sequences, however, the high intrinsic specific absorption rates must be considered in clinical situations. This study introduces a fast BS B1+ mapping method based on a SE‐BURST sequence (BS‐SE‐BURST). With SE‐BURST sequences, multiple low‐magnitude excitation pulses are applied prior to the refocusing pulse. Thus, multiple and different phase‐encoded echoes can be acquired per excitation cycle. Compared with a SE sequence, this excitation strategy results in a similar signal‐to‐noise ratio (SNR) per unit time but with reduced specific absorption rate. The proposed BS‐SE‐BURST sequence was implemented on a conventional 3 T whole body MRI scanner and applied successfully. Magn Reson Med, 2012.