Multispectral fluorescence lifetime imaging system for intravascular diagnostics with ultrasound guidance: in vivo validation in swine arteries

Fluorescence lifetime technique has demonstrated potential for analysis of atherosclerotic lesions and for complementing existing intravascular imaging modalities such as intravascular ultrasound (IVUS) in identifying lesions at high risk of rupture. This study presents a multimodal catheter system integrating a 40 MHz commercial IVUS and fluorescence lifetime imaging (FLIm) using fast helical motion scanning (400 rpm, 0.75 mm/s), able to acquire in vivo in pulsatile blood flow the autofluorescence emission of arterial vessels with high precision (5.08 ± 0.26 ns mean average lifetime over 13 scans). Co‐registered FLIm and IVUS data allowed 3D visualization of both biochemical and morphological vessel properties. Current study supports the development of clinically compatible intravascular diagnostic system integrating FLIm and demonstrates, to our knowledge, the first in vivo intravascular application of a fluorescence lifetime imaging technique. (© 2014 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Strain measurement in coronary arteries using intravascular ultrasound and deformable images

Atherosclerotic plaque rupture is responsible for the majority of myocardial infarctions and acute coronary syndromes. Rupture is initiated by mechanical failure of the plaque cap, and thus study of the deformation of the plaque in the artery can elucidate the events that lead to myocardial infarction. Intravascular ultrasound (IVUS) provides high resolution in vitro and in vivo cross-sectional images of blood vessels. To extract the deformation field from sequences of IVUS images, a registration process must be performed to correlate material points between image pairs. The objective of this study was to determine the efficacy of an image registration technique termed Warping to determine strains in plaques and coronary arteries from paired IVUS images representing two different states of deformation. The Warping technique uses pointwise differences in pixel intensities between image pairs to generate a distributed body force that acts to deform a finite element model. The strain distribution estimated by image-based Warping showed excellent agreement with a known forward finite element solution, representing the gold standard, from which the displaced image was created. The Warping technique had a low sensitivity to changes in material parameters or material model and had a low dependency on the noise present in the images. The Warping analysis was also able to produce accurate strain distributions when the constitutive model used for the Warping analysis and the forward analysis was different. The results of this study demonstrate that Warping in conjunction with in vivo IVUS imaging will determine the change in the strain distribution resulting from physiological loading and may be useful as a diagnostic tool for predicting the likelihood of plaque rupture through the determination of the relative stiffness of the plaque constituents.     

The role of intravascular ultrasound imaging in vascular brachytherapy

Intracoronary brachytherapy has recently emerged as a new therapy to prevent restenosis. Initial experimental work was achieved in animal models and the results were assessed by histomorphometry. Initial clinical trials used angiography to guide dosimetry and to assess efficacy. Intravascular ultrasound (IVUS) permits tomographic examination of the vessel wall, elucidating the true morphology of the lumen and transmural components, which cannot be investigated on the lumenogram obtained by angiography. This paper reviews the use of IVUS in the clinical studies of brachytherapy conducted to date. IVUS allows clinicians to make a thorough assessment of the remodeling of the vessel and appears to have a major role to play in facilitating understanding of the underlying mechanisms of action in this emerging field. The authors propose that state-of-the-art IVUS techniques should be employed to further knowledge of the mechanisms of action of brachytherapy in atherosclerotic human coronary arteries.     

Inducing occlusion effect in Y-shaped vessels using high-intensity focused ultrasound: finite element analysis and phantom validation

High-intensity focused ultrasound (HIFU) surgery offers a truly non-invasive treatment method with no skin incision, but precise targeting of tumour tissues for thermotherapy. Clinical experience reveals that the efficacy of tumour destruction not only involves in coagulating necrosis, but also involves in damaging the tumour vessels, which play an important role in tumour progression. These vessels take the elevated temperature away by perfusion, resulting in uncertainty of the occlusion effect during HIFU treatment. In this study, a Y-shaped vessel model comprising common and tumour vessels and an indirect fabrication method are proposed. The physical properties of the fabricated vessel phantom are measured and compared with human tissue. Simulation is performed using finite element modelling according to the tissue parameter, perfusion rate of the tumour vessel and treatment parameters including power intensity and exposure duration. The phantom experiments are carried out with perfusion of egg white to validate the threshold time prediction obtained from the simulation results. Our findings reveal that the threshold time obtained from experiments is consistent with the simulated one.     

Automated image analysis for array hybridization experiments.

MOTIVATION: Image analysis is a major part of data evaluation for array hybridization experiments in molecular biology. The program presented here is designed to analyze automatically images from hybridization experiments with various arrangements: different kinds of probes (oligonucleotides or complex probes), different supports (nylon filters or glass slides), different labeling of probes (radioactively or fluorescently). The program is currently applied to oligonucleotide fingerprinting projects and complex hybridizations. The only precondition for the use of the program is that the targets are arrayed in a grid, which can be approximately transformed to an orthogonal equidistant grid by a projective mapping. RESULTS: We demonstrate that our program can cope with the following problems: global distortion of the grid, missing of grid nodes, local deviation of the spot from its specified grid position. This is checked by different quality measures. The image analysis of oligonucleotide fingerprint experiments on an entire genetic library is used, in clustering procedures, to group related clones together. The results show that the program yields automatically generated high quality input data for follow up analysis such as clustering procedures. AVAILABILITY: The executable files will be available upon request for academics.     

Vasa vasorum and molecular imaging of atherosclerotic plaques using nonlinear contrast intravascular ultrasound

There is increasing evidence that presence and location of neovascular vasa vasorum play an important role in atherosclerotic plaque pathogenesis and stability. This paper describes a method to detect vasa vasorum with high contrast and high spatial resolution. It uses second harmonic or subharmonic intravascular ultrasound, in combination with ultrasound contrast agents. The same technology in combination with targeted contrast agents is suited for molecular imaging. The potential for vasa vasorum imaging is illustrated using an atherosclerotic animal model and the potential for molecular imaging is illustrated using phantom experiments.     

Strain imaging of the lateral collateral ligament using high frequency and conventional ultrasound imaging: An ex-vivo comparison

Recent first attempts of in situ ultrasound strain imaging in collateral ligaments encountered a number of challenges and illustrated a clear need for additional studies and more thorough validation of the available strain imaging methods. Therefore, in this study we experimentally validated ultrasound strain measurements of ex vivo human lateral collateral ligaments in an axial loading condition. Moreover, the use of high frequency ultrasound (>20 MHz) for strain measurement was explored and its performance compared to conventional ultrasound. The ligaments were stretched up to 5% strain and ultrasound measurements were compared to surface strain measurements from optical digital image correlation (DIC) techniques. The results show good correlations between ultrasound based and DIC based strain measures with R² values of 0.71 and 0.93 for high frequency and conventional ultrasound, subsequently. The performance of conventional ultrasound was significantly higher compared to high frequency ultrasound strain imaging, as the high frequency based method seemed more prone to errors. This study demonstrates that ultrasound strain imaging is feasible in ex vivo lateral collateral ligaments, which are relatively small structures. Additional studies should be designed for a more informed assessment of optimal in vivo strain measurements in collateral knee ligaments.     

Finding coexpressed genes in counts-based data: an improved measure with validation experiments

MOTIVATION: Expressed sequence tag (EST) data reflects variation in gene expression, but previous methods for finding coexpressed genes in EST data are subject to bias and vastly overstate the statistical significance of putatively coexpressed genes. RESULTS: We introduce a new method (LNP) that reports reasonable p-values and also detects more biological relationships in human dbEST than do previous methods. In simulations with human dbEST library sizes, previous methods report p-values as low as 10(-30) on 1/1000 uncorrelated pairs, while LNP reports significance correctly. We validate the analysis on real human genes by comparing coexpressed pairs to gene ontology annotations and find that LNP is more sensitive than the three previous methods. We also find a small but statistically significant level of coexpression between interacting proteins relative to randomized controls. The LNP method is based on a log-normal prior on the distribution of expression levels.     

3D dosimetric validation of ultrasound-guided radiotherapy with a dynamically deformable abdominal phantom

ObjectivesThe purpose of this study was to dosimetrically benchmark gel dosimetry measurements in a dynamically deformable abdominal phantom for intrafraction image guidance through a multi-dosimeter comparison. Once benchmarked, the study aimed to perform a proof-of-principle study for validation measurements of an ultrasound image-guided radiotherapy delivery system.MethodsThe phantom was dosimetrically benchmarked by delivering a liver VMAT plan and measuring the 3D dose distribution with DEFGEL dosimeters. Measured doses were compared to the treatment planning system and measurements acquired with radiochromic film and an ion chamber. The ultrasound image guidance validation was performed for a hands-free ultrasound transducer for the tracking of liver motion during treatment.ResultsGel dosimeters were compared to the TPS and film measurements, showing good qualitative dose distribution matches, low γ values through most of the high dose region, and average 3%/5 mm γ-analysis pass rates of 99.2%(0.8%) and 90.1%(0.8%), respectively. Gel dosimeter measurements matched ion chamber measurements within 3%. The image guidance validation study showed the measurement of the treatment delivery improvements due to the inclusion of the ultrasound image guidance system. Good qualitative matching of dose distributions and improvements of the γ-analysis results were observed for the ultrasound-gated dosimeter compared to the ungated dosimeter.ConclusionsDEFGEL dosimeters in phantom showed good agreement with the planned dose and other dosimeters for dosimetric benchmarking. Ultrasound image guidance validation measurements showed good proof-of-principle of the utility of the phantom system as a method of validating ultrasound-based image guidance systems and potentially other image guidance methods.     

Molecular imaging with targeted contrast ultrasound

Molecular imaging with contrast ultrasound relies on the detection of targeted microbubbles or other acoustically active nanoparticles. These microbubbles are retained in diseased tissue where they produce an acoustic signal because of their resonant properties in the ultrasound field. Targeting is accomplished either through manipulating the chemical properties of the microbubble shell or through conjugation of disease-specific ligands for the targeted molecule to the microbubble surface. As microbubbles cannot leave the intravascular space, the disease process must be characterized by molecular changes in the vascular compartment to be imaged. Inflammation, angiogenesis and thrombus formation are central pathophysiologic processes in many disease states and produce phenotypic changes in the vascular compartment. Thus, targeted contrast ultrasound in the future could aid in the diagnosis of such diverse diseases as atherosclerosis, transplant rejection and tumor-related angiogenesis.     

Acute coronary closure after stenting: a lesson from intravascular ultrasound

Intravascular ultrasound (IVUS) is an imaging modality more sensitive for defining suboptimal stent deployment issues compared with conventional angiographic imaging. We report on a patient with unstable angina who underwent stent deployment using IVUS guidance. Despite an acceptable angiographic result, intrastent thrombus, incomplete expansion and an edge/marginal dissection were seen by IVUS examination. Both the incomplete expansion and the intrastent thrombus were successfully treated. The dissection, however, was not treated in the catheterization laboratory. This mechanical injury pattern at the stent edge led to vessel occlusion 4 h post-procedure.     

Abnormal pulmonary artery stiffness in pulmonary arterial hypertension: in vivo study with intravascular ultrasound

Background

There is increasing recognition that pulmonary artery stiffness is an important determinant of right ventricular (RV) afterload in pulmonary arterial hypertension (PAH). We used intravascular ultrasound (IVUS) to evaluate the mechanical properties of the elastic pulmonary arteries (PA) in subjects with PAH, and assessed the effects of PAH-specific therapy on indices of arterial stiffness.

Method

Using IVUS and simultaneous right heart catheterisation, 20 pulmonary segments in 8 PAH subjects and 12 pulmonary segments in 8 controls were studied to determine their compliance, distensibility, elastic modulus and stiffness index β. PAH subjects underwent repeat IVUS examinations after 6-months of bosentan therapy.

Results

At baseline, PAH subjects demonstrated greater stiffness in all measured indices compared to controls: compliance (1.50±0.11×10^–2 mm^2/mmHg vs 4.49±0.43×10^–2 mm^2/mmHg, p<0.0001), distensibility (0.32±0.03%/mmHg vs 1.18±0.13%/mmHg, p<0.0001), elastic modulus (720±64 mmHg vs 198±19 mmHg, p<0.0001), and stiffness index β (15.0±1.4 vs 11.0±0.7, p = 0.046). Strong inverse exponential associations existed between mean pulmonary artery pressure and compliance (r² = 0.82, p<0.0001), and also between mean PAP and distensibility (r² = 0.79, p = 0.002). Bosentan therapy, for 6-months, was not associated with any significant changes in all indices of PA stiffness.

Conclusion

Increased stiffness occurs in the proximal elastic PA in patients with PAH and contributes to the pathogenesis RV failure. Bosentan therapy may not be effective at improving PA stiffness.     

Magnetic particle imaging for artifact-free imaging of intracranial flow diverter stents: A phantom study

PurposeMagnetic Particle Imaging (MPI) is a new, background- and radiation-free tomographic imaging method that enables near real-time imaging of superparamagnetic iron-oxide nanoparticles (SPIONs) with high temporal and spatial resolution. This phantom study aims to investigate the potential of MPI for visualization of the stent lumen in intracranial flow diverters (FD).MethodsNitinol FD of different dimensions (outer diameter: 3.5 mm, 4.0 mm, 5.5 mm; total length: 22–40 mm) were scanned in vascular phantoms in a custom-built MPI scanner (in-plane resolution: ~ 2 mm, field of view: 65 mm length, 29 mm diameter). Phantoms were filled with diluted (1:50) SPION tracer agent Ferucarbotran (10 µmol (Fe)/ml; NaCL). Each phantom was measured in 32 different projections (overall acquisition time per image: 3200 ms, 5 averages). After image reconstruction from raw data, two radiologists assessed image quality using a 5-point Likert scale. The signal intensity profile was measured using a semi-automatic evaluation tool.ResultsMPI visualized the lumen of all FD without relevant differences between the stented vessel phantom and the reference phantom. At 3.5 mm image quality was slightly inferior to the larger diameters. The FD themselves neither generated an MPI signal nor did they lead to relevant imaging artifacts. Ratings of both radiologists showed no significant difference, interrater reliability was good (ICC 0.84). A quantitative evaluation of the signal intensity profile did not reveal any significant differences (p > 0.05) either.ConclusionMPI visualizes the lumen of nitinol FD stents in vessel phantoms without relevant stent-induced artifacts.     

Imaging of left main trauma during diagnostic cardiac catheterization with intravascular ultrasound

In this case report the occurrence of a catheter-induced coronary artery dissection is described. In our patient, angiography showed a mushroom-shaped exudate above the left main coronary artery. Intravascular ultrasound revealed a circular dissection with a huge false lumen connected to the true lumen by a small intimal tear. A brief review of the literature on catheter-induced coronary dissection is included. We believe that this case report provides a good illustration of the need for careful reviewing of indications for angiography. Although procedural risks are low, angiography remains an invasive diagnostic test with the potential to cause severe complications.     

Functional imaging of plants by magnetic resonance experiments.

Microimaging based on magnetic resonance is an experimental technique that can provide a unique view of a variety of plant physiological processes. Particularly interesting applications include investigations of water movement and spatially resolved studies of the transport and accumulation of labelled molecules in intact plant tissue. Some of the fundamental principles of nuclear and electron magnetic resonance microimaging are explained here and the potential of these techniques is shown using several representative examples.     

Calibration and validation of confocal spectral imaging systems.

BACKGROUND: Confocal spectral imaging (CSI) microscopic systems currently on the market delineate multiple fluorescent proteins, labels, or dyes within biological specimens by performing spectral characterizations. However, some CSI systems have been found to present inconsistent spectral profiles of reference spectra within a particular system and between related and unrelated instruments. This variability confirms that there is a need for a standardized, objective calibration and validation protocol. METHODS: Our protocol uses an inexpensive multi-ion discharge lamp (MIDL) that contains Hg(+), Ar(+), and inorganic fluorophores that emit distinct, stable, spectral features in place of a sample. We derived reference spectra from the MIDL data to accurately predict the spectral resolution, ratio of wavelength to wavelength, contrast, and aliasing parameters of any CSI system. We were also able to predict and confirm the influence of pinhole diameter on spectral profiles. RESULTS: Using this simulation, we determined that there was good agreement between observed and theoretical expectations, thus enabling us to identify malfunctioning subsystems. We examined eight CSI systems and one nonconfocal spectral system, all of which displayed spectral inconsistencies. No instrument met its optimal performance expectations. In two systems, we established the need for factory realignment that had not been otherwise recognized. CONCLUSIONS: We found that using a primary light source that emits an absolute standard "reference spectrum" enabled us to diagnose instrumental errors and measure accuracy and reproducibility under normalized conditions. With this information, a CSI operator can determine whether a CSI system is working optimally and make objective comparisons with the performance of other CSI systems. We determined that, if CSI systems were standardized to produce the same spectral profile of a MIDL lamp, researchers could be confident that the same experimental findings would be obtained on any CSI system.