Langendorff perfused hearts have been frequently studied in recent years using optical fluorescence imaging. This in vitro approach, which enables the heart to continue beating after extraction from the body of the animal, allows investigation of physiological functions with relative simplicity compared to in vivo setups. For example, when combined with voltage- and calcium- sensitive dyes, optical mapping of transmembrane potential, calcium transients, and other parameters can lead to a better understanding of cardiac mechanisms underlying heart failures and diseases. However, biomedical optical imaging is fundamentally limited to superficial investigations due to light scattering in tissues, restricting mapping to the heart surface only. The ability to visualize the heart septum would be important for comprehensive cardiac research. While 3D ultrasound can offer imaging of the entire heart, it can only provide mechanical contrast and the spatio-temporal resolution is also insufficient for imaging the heart in 3D on a beat-by-beat basis. Herein, we investigate on the capabilities of optoacoustic tomographic imaging of the Langendorff heart. The heart isolation method allows direct imaging without the presence of surrounding tissues and reduced blood content, significantly improving the penetration depth as well as image quality. The imaging system can acquire 3D images of the heart with optical contrast at an imaging rate of 100 Hz and 150 µm resolution. This enables capturing beat-by-beat heart motion with temporal resolution of 33 sampling instances per heartbeat. The high spatial resolution also allows identifying important internal heart features, including the septum, valves, cordae tendineae, and papillary muscles.
The atrial pacemaker complex is responsible for the initiation and early propagation of cardiac impulses. Optical coherence tomography (OCT), a nondestructive imaging modality with spatial resolutions of ∼ 1 to 15 μm, can be used to identify unique fiber orientation patterns in this region of the heart. Functionally characterized canine sinoatrial nodes (SAN) (n = 7) were imaged using OCT up to ∼ 1 mm below the endocardial tissue surface. OCT images were directly compared to their corresponding histological sections. Fiber orientation patterns unique to the crista terminalis (CT), SAN, and surrounding atrial myocardium were identified with dominant average fiber angles of 89±12 deg, 110±16 deg, and 95±35 deg, respectively. Both the CT and surrounding atrial myocardium displayed predominantly unidirectionally based fiber orientation patterns within each specimen, whereas the SAN displayed an increased amount of fiber disarray manifested quantitatively as a significantly greater standard deviation in fiber angle distribution within specimens [33±7 deg versus 23±5 deg, atrium (p = 0.02); 18±3 deg, CT (p = 0.0003)]. We also identified unique, local patterns of fiber orientation specific to the functionally characterized block zone. We demonstrate the ability of OCT in detecting components of the atrial pacemaker complex which are intimately involved in both normal and abnormal cardiac conduction.
Optical coherence tomography (OCT) allows for the visualization of micron-scale structures within nontransparent biological tissues. For the first time, we demonstrate the use of OCT in identifying components of the cardiac conduction system and other structures in the explanted human heart. Reconstructions of cardiac structures up to 2 mm below the tissue surface were achieved and validated with Masson Trichrome histology in atrial, ventricular, sinoatrial nodal, and atrioventricular nodal preparations. The high spatial resolution of OCT provides visualization of cardiac fibers within the myocardium, as well as elements of the cardiac conduction system; however, a limiting factor remains its depth penetration, demonstrated to be ~2 mm in cardiac tissues. Despite its currently limited imaging depth, the use of OCT to identify the structural determinants of both normal and abnormal function in the intact human heart is critical in its development as a potential aid to intracardiac arrhythmia diagnosis and therapy.
The development of systems physiology is hampered by the limited ability to relate tissue structure and function in intact organs in vivo or in vitro. Here, we show the application of a bimodal biophotonic imaging approach that employs optical coherence tomography and fluorescent imaging to investigate the structure-function relationship at the tissue level in the heart. Reconstruction of cardiac excitation and structure was limited by the depth penetration of bimodal imaging to ~2 mm in atrial tissue, and ~1 mm in ventricular myocardium. The subcellular resolution of optical coherence tomography clearly demonstrated that microscopic fiber orientation governs the pattern of wave propagation in functionally characterized rabbit sinoatrial and atrioventricular nodal preparations and revealed structural heterogeneities contributing to ventricular arrhythmias. The combination of this bimodal biophotonic imaging approach with histology and/or immunohistochemistry can span multiple scales of resolution for the investigation of the molecular and structural determinants of intact tissue physiology.
Heterogeneity in cardiac tissue microstructure is a potential mechanism for the generation and maintenance of arrhythmias. Abnormal changes in fiber orientation increase the likelihood of arrhythmia. We present optical coherence tomography (OCT) as a method to image myofibers in excised intact heart preparations. Three-dimensional (3-D) image sets were gathered from the rabbit right ventricular free wall (RVFW) using a microscope-integrated OCT system. An automated algorithm for fiber orientation quantification in the plane parallel to the wall surface was developed. The algorithm was validated by comparison with manual measurements. Quantifying fiber orientation in the plane parallel to the wall surface from OCT images can be used to help understand the conduction system of the specific sample being imaged.
KEYWORDS: Heart, Personal digital assistants, Signal to noise ratio, Panoramic photography, Imaging systems, Action potentials, CCD cameras, Luminescence, 3D image processing, Light emitting diodes
Cardiac fluorescent optical imaging provides the unique opportunity to investigate the dynamics of propagating electrical waves during ventricular arrhythmias and the termination of arrhythmias by strong electric shocks. Panoramic imaging systems using charge-coupled device (CCD) cameras as the photodetector have been developed to overcome the inability to monitor electrical activity from the entire cardiac surface. Photodiode arrays (PDAs) are known to have higher temporal resolution and signal quality, but lower spatial resolution compared to CCD cameras. We construct a panoramic imaging system with three PDAs and image Langendorff perfused rabbit hearts (n=18) during normal sinus rhythm, epicardial pacing, and arrhythmias. The recorded spatiotemporal dynamics of electrical activity is texture mapped onto a reconstructed 3-D geometrical heart model specific to each heart studied. The PDA-based system provides sufficient spatial resolution (1.72 mm without interpolation) for the study of wavefront propagation in the rabbit heart. The reconstructed 3-D electrical activity provides us with a powerful tool to investigate the fundamental mechanisms of arrhythmia maintenance and termination.
Computed tomography (CT), ultrasound, and magnetic resonance imaging have been used to image and diagnose diseases of the human heart. By gating the acquisition of the images to the heart cycle (gated imaging), these modalities enable one to produce 3D images of the heart without significant motion artifact and to more accurately calculate various parameters such as ejection fractions [1-3]. Unfortunately, these imaging modalities give inadequate resolution when investigating embryonic development in animal models. Defects in developmental mechanisms during embryogenesis have long been thought to result in congenital cardiac anomalies. Our understanding of normal mechanisms of heart development and how abnormalities can lead to defects has been hampered by our inability to detect anatomic and physiologic changes in these small (<2mm) organs. Optical coherence tomography (OCT) has made it possible to visualize internal structures of the living embryonic heart with high-resolution in two- and threedimensions. OCT offers higher resolution than ultrasound (30 um axial, 90 um lateral) and magnetic resonance microscopy (25 um axial, 31 um lateral) [4, 5], with greater depth penetration over confocal microscopy (200 um). Optical coherence tomography (OCT) uses back reflected light from a sample to create an image with axial resolutions ranging from 2-15 um, while penetrating 1-2 mm in depth [6]. In the past, OCT groups estimated ejection fractions using 2D images in a Xenopus laevis [7], created 3D renderings of chick embryo hearts [8], and used a gated reconstruction technique to produce 2D Doppler OCT image of an in vivo Xenopus laevis heart [9].
In this paper we present a gated imaging system that allowed us to produce a 16-frame 3D movie of a beating chick embryo heart. The heart was excised from a day two (stage 13) chicken embryo and electrically paced at 1 Hz. We acquired 2D images (B-scans) in 62.5 ms, which provides enough temporal resolution to distinguish end-contraction from end-relaxation. After acquiring the image set, we were able to measure the ejection fraction.
KEYWORDS: Optical coherence tomography, Heart, 3D image processing, Control systems, Tissues, In vivo imaging, Image resolution, Photography, Visualization, Frame grabbers
Using Optical Coherence Tomography (OCT), an emerging imaging modality, we have produced both 3D and 4D images of cardiac architecture. We captured 3D images of rabbit Purkinje fiber networks and we also created a 4D representation of a beating stage 28 chicken embryo heart. For the 4D reconstruction, we generated a movie by employing a gated reconstruction technique.
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