Coronary atherosclerosis is caused by plaque build-up, with lipids playing a pivotal role in its progression. However, lipid composition and distribution within coronary atherosclerosis remain unknown. This study aims to characterize lipids and investigate differences in lipid composition across disease stages to aid in the understanding of disease progression. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) was used to visualize lipid distributions in coronary artery sections (n ¼ 17) from hypercholesterolemic swine. We performed histology on consecutive sections to classify the artery segments and to investigate colocalization between lipids and histological regions of interest in advanced plaque, including necrotic core and inflammatory cells. Segments were classified as healthy (n ¼ 6), mild (n ¼ 6), and advanced disease (n ¼ 5) artery segments. Multivariate data analysis was employed to find differences in lipid composition between the segment types, and the lipids' spatial distribution was investigated using non-negative matrix factorization (NMF). Through this process, MALDI-MSI detected 473 lipid-related features. NMF clustering described three components in positive ionization mode: triacylglycerides (TAG), phosphatidylcholines (PC), and cholesterol species. In negative ionization mode, two components were identified: one driven by phosphatidylinositol(PI)(38:4), and one driven by ceramidephosphoethanolamine(36:1). Multivariate data analysis showed the association between advanced disease and specific lipid signatures like PC(O-40:5) and cholesterylester(CE)(18:2). Ether-linked phospholipids and LysoPC species were found to colocalize with necrotic core, and mostly CE, ceramide, and PI species colocalized with inflammatory cells. This study, therefore, uncovers distinct lipid signatures correlated with plaque development and their colocalization with necrotic core and inflammatory cells, enhancing our understanding of coronary atherosclerosis progression.

Due to the shortage of kidneys donated for transplantation, surgeons are forced to use the organs with an elevated risk of poor function or even failure. Although the existing methods for pre-transplant quality evaluation have been validated over decades in population cohort studies across the world, new methods are needed as long as delayed graft function or failure in a kidney transplant occurs. In this study, we explored the potential of utilizing photoacoustic (PA) imaging during normothermic machine perfusion (NMP) as a means of evaluating kidney quality. We closely monitored twenty-two porcine kidneys using 3D PA imaging during a two-hour NMP session. Based on biochemical analyses of perfusate and produced urine, the kidneys were categorized into ‘non-functional’ and ‘functional’ groups. Our primary focus was to quantify oxygenation (sO₂) within the kidney cortical layer of depths 2 mm, 4 mm, and 6 mm using two-wavelength PA imaging. Next, receiver operating characteristic (ROC) analysis was performed to determine an optimal cortical layer depth and time point for the quantification of sO₂ to discriminate between functional and non-functional organs. Finally, for each depth, we assessed the correlation between sO₂ and creatinine clearance (CrCl), oxygen consumption (VO₂), and renal blood flow (RBF). We found that hypoxia of the renal cortex is associated with poor renal function. In addition, the determination of sO₂ within the 2 mm depth of the renal cortex after 30 min of NMP effectively distinguishes between functional and non-functional kidneys. The non-functional kidneys can be detected with the sensitivity and specificity of 80% and 85% respectively, using the cut-off point of sO₂ < 39%. Oxygenation significantly correlates with RBF and VO₂ in all kidneys. In functional kidneys, sO₂ correlated with CrCl, which is not the case for non-functional kidneys. We conclude that the presented technique has a high potential for supporting organ selection for kidney transplantation.

Photothermal optical coherence microscopy (PT-OCM) combines the high-resolution, label-free morphological imaging of OCM with the ability to discriminate tissue composition through phase-sensitive photothermal imaging. In this study, we perform 2D imaging of human carotid endarterectomies to spectrally determine lipid distribution, with verification via histologically stained samples. The structural information from OCM is combined with the spectral information gained from measuring the resulting sample surface displacement from thermoelastic expansion, following light irradiation. PT-OCM is thus demonstrated as a potential tool in the investigation of atherosclerotic plaque lipids, contributing towards the understanding of plaque instability.

Significance: Spectroscopic analysis of optical coherence tomography (OCT) data can yield added information about the sample's chemical composition, along with high-resolution images. Typical commercial OCT systems operate at wavelengths that may not be optimal for identifying lipid-containing samples based on absorption features. Aim: The main aim of this study was to develop a 1200 nm spectroscopic OCT (SOCT) for the classification of lipid-based and water-based samples by extracting the lipid absorption peak at 1210 nm from the OCT data. Approach: We developed a 1200 nm OCT system and implemented a signal processing algorithm that simultaneously retrieves spectroscopic and structural information from the sample. In this study, we validated the performance of our OCT system by imaging weakly scattering phantoms with and without lipid absorption features. An orthogonal projections to latent structures-discriminant analysis (OPLS-DA) model was developed and applied to classify weakly scattering samples based on their absorption features. Results: The OCT system achieved an axial resolution of 7.2 m and a sensitivity of 95 dB. The calibrated OPLS-DA model on weakly scattering samples with lipid and water-based absorption features correctly classified 19/20 validation samples. Conclusions: The 1200 nm SOCT system can discriminate the lipid-containing weakly scattering samples from water-based weakly scattering samples with good predictive ability.

Background and aims: Lipids play an important role in atherosclerotic plaque development and are interesting candidate predictive biomarkers. However, the link between circulating lipids, accumulating lipids in the vessel wall, and plaque destabilization processes in humans remains largely unknown. This study aims to provide new insights into the role of lipids in atherosclerosis using lipidomics and mass spectrometry imaging to investigate lipid signatures in advanced human carotid plaque and plasma samples. Methods: We used lipidomics and desorption electrospray ionization mass spectrometry imaging (DESI-MSI) to investigate lipid signatures of advanced human carotid plaque and plasma obtained from patients who underwent carotid endarterectomy (n = 14 out of 17 whose plaque samples were analyzed by DESI-MSI). Multivariate data analysis and unsupervised clustering were applied to identify lipids that were the most discriminative species between different patterns in plaque and plasma. These patterns were interpreted by quantitative comparison with conventional histology. Results: Lipidomics detected more than 300 lipid species in plasma and plaque, with markedly different relative abundances. DESI-MSI visualized the spatial distribution of 611 lipid-related m/z features in plaques, of which 330 m/z features could be assigned based on exact mass, comparison to the lipidomic data, and high mass resolution MSI. Matching spatial lipid patterns to histological areas of interest revealed several molecular species that were colocalized with pertinent disease processes in plaque including specific sphingomyelin and ceramide species with calcification, phospholipids and free fatty acids with inflammation, and triacylglycerols and phosphatidylinositols with fibrin-rich areas. Conclusions: By comparing lipid species in plaque and plasma, we identified those circulating species that were also prominently present in plaque. Quantitative comparison of lipid spectral patterns with histology revealed the presence of specific lipid species in destabilized plaque areas, corroborating previous in vitro and animal studies.

A dual frequency probe using a multi-layer piezoelectric material is proposed for simultaneous ultrasound and photoacoustic imaging of the carotid artery with a high resolution ultrasound and a high sensitivity photoacoustic image. The probe consists of lead zirconate titanate (PZT) for ultrasound stack and and polyvinylidene difluoride (PVDF) array for photoacoustic signal reception, which is placed on top of the PZT stack. We used 3D finite element analysis to evaluate a quarter of the full aperture of the dual frequency array, having 48 elements diced PZT-5H for ultrasound pulse-echo and 16 elements of 28µm thick, kerfless PVDF for photoacoustic receiving. We showed that considering the PVDF array as the second matching layer of the ultrasound stack minimized its loading effect at the cost of operating in a higher operation frequency of 9.9 MHz. We modeled a design with and without sub-dicing, where sub-dicing and subsequent suppression of lateral modes allows larger elements and thus larger aperture. The -3dB bandwidth of the ultrasound stack with and without sub-dicing are 87% and 75% relative to the center frequencies. We found a transmit sensitivity of 17 kPa/V and 21 kPa/V for those two realizations respectively.

We demonstrate megahertz intravascular Doppler Optical Coherence Tomography (OCT). The OCT system relies on a 1.1 mm diameter motorized catheter and a 1.5 MHz Fourier Domain Mode Locked (FDML) laser. By resolving the phase shift between adjoint A-lines, the flow pattern image can be reconstructed. Imaging experiments were carried out in swine coronary artery in vitro at a speed of 600 frames/s. The MHz sweep rate not only allows us to investigate flow velocity of up to 37.5 cm/s without phase-unwrapping, but also enables dynamic flow imaging at high frame rate.

We present a form of acoustic microscopy, called Structured Ultrasound Microscopy (SUM). It creates a volumetric image by recording reflected echoes of ultrasound waves with a structured phase front using a moving single-element transducer and computational reconstruction. A priori knowledge of the acoustic field produced by the single element allows us to relate the received echoes to a 3D scatter map within the acoustic beam itself, leading to an isotropic resolution at all depths. An aberration mask in front of the acoustic element imposes the phase structure, broadening the beam and breaking the spatial coherence between different voxels at equal acoustic propagation delay, increasing the uniqueness of the reconstruction. By translating the transducer across the 3D volume, we synthetically enlarge the imaging aperture by using multiple overlapping and spatially sparsely sampled measurements to solve for the entire image. In this paper, we explain the SUM technique and demonstrate microscopic imaging at 20 MHz of a 2.3 × 2.3 × 1.2 mm object in water, with an isotropic resolution below 100 μm. The proposed approach allows for wide-field 3D imaging at isotropic microscopic resolution using a small unfocused ultrasound sensor and multiple spatially sparsely sampled measurements. This technique may find applications in many other fields where space is constrained, device simplicity is desired, and wide-field isotropic high-resolution imaging is required.

This paper presents an area- and power-efficient application-specified integrated circuit (ASIC) for 3-D forward-looking intravascular ultrasound imaging. The ASIC is intended to be mounted at the tip of a catheter, and has a circular active area with a diameter of 1.5 mm on the top of which a 2-D array of piezoelectric transducer elements is integrated. It requires only four micro-coaxial cables to interface 64 receive (RX) elements and 16 transmit (TX) elements with an imaging system. To do so, it routes high-voltage (HV) pulses generated by the system to selected TX elements using compact HV switch circuits, digitizes the resulting echo signal received by a selected RX element locally, and employs an energy-efficient load-modulation datalink to return the digitized echo signal to the system in a robust manner. A multi-functional command line provides the required sampling clock, configuration data, and supply voltage for the HV switches. The ASIC has been realized in a 0.18-&#x03BC;m HV CMOS technology and consumes only 9.1 mW. Electrical measurements show 28-V HV switching and RX digitization with a 16-MHz bandwidth and 53-dB dynamic range. Acoustical measurements demonstrate successful pulse transmission and reception. Finally, a 3-D ultrasound image of a three-needle phantom is generated to demonstrate the imaging capability.

Catheter-based radiofrequency ablation for atrial fibrillation has long-term success in 60-70% of cases. A better assessment of lesion quality, depth, and continuity could improve the procedure’s outcome. We investigate here photoacoustic contrast between ablated and healthy atrial-wall tissue in vitro in wavelengths spanning from 410 nm to 1000 nm. We studied single-and multi-wavelength imaging of ablation lesions and we demonstrate that a two-wavelength technique yields precise detection of lesions, achieving a diagnostic accuracy of 97%. We compare this with a best single-wavelength (640 nm) analysis that correctly identifies 82% of lesions. We discuss the origin of relevant spectroscopic features and perspectives for translation to clinical imaging.

Intravascular ultrasound is an imaging modality used to visualize atherosclerosis from within the inner lumen of human arteries. Complex lesions like chronic total occlusions require forward-looking intravascular ultrasound (FL-IVUS), instead of the conventional side-looking geometry. Volumetric imaging can be achieved with 2D array transducers, which present major challenges in reducing cable count and device integration. In this work we present an 80-element lead zirconium titanate (PZT) matrix ultrasound transducer for FL-IVUS imaging with a front-end application-specific integrated circuit (ASIC) requiring only 4 cables. After investigating optimal transducer designs we fabricated the matrix transducer consisting of 16 transmit (TX) and 64 receive (RX) elements arranged on top of an ASIC having an outer diameter of 1.5 mm and a central hole of 0.5 mm for a guidewire. We modeled the transducer using finite element analysis and compared the simulation results to the values obtained through acoustic measurements. The TX elements showed uniform behavior with a center frequency of 14 MHz, a -3 dB bandwidth of 44 &#x0025; and a transmit sensitivity of 0.4 kPa/V at 6 mm. The RX elements showed center frequency and bandwidth similar to the TX elements, with an estimated receive sensitivity of 3.7 &#x03BC;V/Pa. We successfully acquired a 3D FL image of three spherical reflectors in water using delay-and-sum beamforming and the coherence factor method. Full synthetic aperture acquisition can be achieved with frame rates on the order of 100 Hz. The acoustic characterization and the initial imaging results show the potential of the proposed transducer to achieve 3D FL-IVUS imaging.

Lipid deposition can be assessed with combined intravascular photoacoustic/ultrasound (IVPA/US) imaging. To date, the clinical translation of IVPA/US imaging has been stalled by a low imaging speed and catheter complexity. In this paper, we demonstrate imaging of lipid targets in swine coronary arteries in vivo, at a clinically useful frame rate of 20 s⁻¹. We confirmed image contrast for atherosclerotic plaque in human samples ex vivo. The system is on a mobile platform and provides real-time data visualization during acquisition. We achieved an IVPA signal-to-noise ratio of 20 dB. These data show that clinical translation of IVPA is possible in principle.

Photoacoustic imaging couples the chemical specificity of optical absorption with the viewing depth of ultrasound. Systems based on linear array transducers have the versatility to be applied in various (pre-) clinical scenarios but face a trade-off between viewing depth and image resolution depending on transducer frequency and aperture. We propose here a method to disentangle, with precision, small, closely spaced targets with optical spectral contrast, without impairing the imaging depth. Photoacoustic data sets were recorded at two different optical wavelengths. We accurately recovered object separation distances (mean error = 2.3% ô 6%) from the phase difference between signals across the array, down to a spacing of 1/20th of the system's beam-formed lateral resolution. The proposed method may enable the translation of super-resolution microscopy to deep tissue imaging.

Lipid-core atherosclerotic plaques are associated with disease progression, procedural complications, and cardiac events. Coronary plaque lipid can be quantified in optical coherence tomography (OCT) pullbacks by measurement of lipid arcs and lipid lengths; parameters frequently used in clinical research, but labor intensive and subjective to analyse. In this study, we investigated the ability of quantitative attenuation, derived from intravascular OCT, to detect plaque lipid. Lipid cores are associated with a high attenuation coefficient. We compared the index of plaque attenuation (IPA), a local quantitative measure of attenuation, to the manually measured lipid score (arc and length) on OCT images, and to the plaque characterization ex-vivo. We confirmed a correlation between the IPA and lipid scores (r² > 0.7). Comparison to histology shows that high attenuation is associated with fibroatheroma, but also with macrophage presence. IPA is a robust, reproducible, and user-independent measure that facilitates quantification of coronary lipid, a potential tool in clinical research and in guiding percutaneous coronary intervention.

Purpose: Quantitative and automatic analysis of intracoronary optical coherence tomography images is useful and time-saving to assess cardiovascular risk in the clinical arena. Methods: First, the interfaces of the intima, media, and adventitia layers are segmented, by means of an original front propagation scheme, running in a 4D multi-parametric space, to simultaneously extract three non-crossing contours in the initial cross-sectional image. Second, information resulting from the tentative contours is exploited by a machine learning approach to identify healthy and diseased regions of the arterial wall. The framework is fully automatic. Results: The method was applied to 40 patients from two different medical centers. The framework was trained on 140 images and validated on 260 other images. For the contour segmentation method, the average segmentation errors were 29±46μm for the intima–media interface, 30±50μm for the media–adventitia interface, and 50±64μm for the adventitia–periadventitia interface. The classification method demonstrated a good accuracy, with a median Dice coefficient equal to 0.93 and an interquartile range of (0.78–0.98). Conclusion: The proposed framework demonstrated promising offline performances and could potentially be translated into a reliable tool for various clinical applications, such as quantification of tissue layer thickness and global summarization of healthy regions in entire pullbacks.

This paper presents a front-end ASIC for forward-looking intravascular ultrasound (IVUS) imaging. The ASIC is intended to be mounted at the tip of a catheter and can interface a total of 80 piezo-electric transducer elements with an imaging systems using only 4 cables, thus significantly reducing the system complexity compared to the prior art. It is capable of switching high-voltage transmit pulses to 16 transmit elements, and capturing the resulting echo signals using 64 multiplexed receive elements. The ASIC digitizes the received signals locally, providing more robust communication than prior analog approaches. Measurements show that the ASIC effectively switches transmit pulses up to 30 V, and digitizes echo signals with a bandwidth of 16 MHz, while consuming only 10 mW. Acoustic measurements in combination with a prototype transducer array demonstrate pulse transmission and reception. Finally, a B-mode image of a needle phantom demonstrates the imaging capability.

The identification of unstable atherosclerotic plaques in the coronary arteries is emerging as an important tool for guiding percutaneous coronary interventions and may enable preventive treatment of such plaques in the future. Assessment of plaque stability requires imaging of both structure and composition. Spectroscopic photoacoustic (sPA) imaging can visualize atherosclerotic plaque composition on the basis of the optical absorption contrast. It is an established fact that the frequency content of the photoacoustic (PA) signal is correlated with structural tissue properties. As PA signals can be weak, it is important to match the transducer bandwidth to the signal frequency content for in vivo imaging. In this ex vivo study on human coronary arteries, we combined sPA imaging and analysis of frequency content of the PA signals. Using a broadband transducer (-3-dB one-way bandwidth of 10-35 MHz) and a 1-mm needle hydrophone (calibrated for 1-20 MHz), we covered a large frequency range of 1-35 MHz for receiving the PA signals. Spectroscopic PA imaging was performed at wavelengths ranging from 1125 to 1275 nm with a step of 2 nm, allowing discrimination between plaque lipids and adventitial tissue. Under sPA imaging guidance, the frequency content of the PA signals from the plaque lipids was quantified. Our data indicate that more than 80% of the PA energy of the coronary plaque lipids lies in the frequency band below 8 MHz. This frequency information can guide the choice of the transducer element used for PA catheter fabrication.

In medical ultrasound transducer design, the geometry of the individual elements is crucial since it affects the vibration mode of each element and its radiation impedance. For a fixed frequency, optimal vibration (i.e., uniform surface motion) can be achieved by designing elements with very small width-to-thickness ratios. However, for optimal radiation impedance (i.e., highest radiated power), the width should be as large as possible. This leads to a contradiction that can be solved by subdicing wide elements. To systematically examine the effect of subdicing on the performance of a 1-D ultrasound transducer array, we applied finite-element simulations. We investigated the influence of subdicing on the radiation impedance, on the time and frequency response, and on the directivity of linear arrays with variable element widths. We also studied the effect of varying the depth of the subdicing cut. The results show that, for elements having a width greater than 0.6 times the wavelength, subdicing improves the performance compared with that of nonsubdiced elements: the emitted pressure may be increased up to a factor of three, the ringing time may be reduced by up to 50%, the bandwidth increased by up to 77%, and the sidelobes reduced by up to 13 dB. Moreover, this simulation study shows that all these improvements can already be achieved by subdicing the elements to a depth of 70% of the total element thickness. Thus, subdicing can improve important transducer parameters and, therefore, help in achieving images with improved signal-to-noise ratio and improved resolution.