Riqiang Lin

Intravascular optical-resolution photoacoustic tomography with a 1.1 mm diameter catheter.

Photoacoustic imaging is an emerging technology that can provide anatomic, functional, and molecular information about biological tissue. Intravascular spectroscopic and molecular photoacoustic imaging can potentially improve the identification of atherosclerotic plaque composition, the detection of inflammation, and ultimately the risk stratification of atherosclerosis. In this study, a first-of-its-kind intravascular optical-resolution photoacoustic tomography (OR-PAT) system with a 1.1 mm diameter catheter is developed, offering optical-diffraction limited transverse resolution as fine as 19.6 μm, ∼10-fold finer than that of conventional intravascular photoacoustic and ultrasonic imaging. To offer complementary imaging information and depth, the system also acquires co-registered intravascular ultrasound images in parallel. Imaging of an iliac stent and a lipid phantom shows that the high resolution and contrast of OR-PAT can enable improved stent implantation guidance and lipid identification. In the future, these capabilities may ultimately improve the diagnosis and interventional treatment of vulnerable atherosclerotic plaques, which are prone to cause thrombotic complications such as myocardial infarction and stroke.

High-speed intravascular spectroscopic photoacoustic imaging at 1000 A-lines per second with a 0.9-mm diameter catheter

Abstract. Intravascular spectroscopic photoacoustic technology can image atherosclerotic plaque composition with high sensitivity and specificity, which is critical for identifying vulnerable plaques. Here, we designed and engineered a catheter of 0.9 mm in diameter for intravascular photoacoustic (IVPA) imaging, smaller than the critical size of 1 mm required for clinical translation. Further, a quasifocusing photoacoustic excitation scheme was developed for the catheter, producing well-detectable IVPA signals from stents and lipids with a laser energy as low as ∼30  μJ/pulse. As a result, this design enabled the use of a low-energy, high-repetition rate, ns-pulsed optical parametric oscillator laser for high-speed spectroscopic IVPA imaging at both the 1.2-μm and 1.7-μm spectral bands for lipid detection. Specifically, for each wavelength, a 1-kHz IVPA A-line rate was achieved, ∼100-fold faster than previously reported IVPA systems offering a similar wavelength tuning range. Using the system, spectroscopic IVPA imaging of peri-adventitial adipose tissue from a porcine aorta segment was demonstrated. The significantly improved imaging speed, together with the reduced catheter size and multiwavelength spectroscopic imaging ability, suggests that the developed high-speed IVPA technology is of great potential to be further translated for in vivo applications.

Blind-deconvolution optical-resolution photoacoustic microscopy in vivo

Optical-resolution photoacoustic microscopy (OR-PAM) is becoming a vital tool for studying the microcirculation system in vivo. By increasing the numerical aperture of optical focusing, the lateral resolution of OR-PAM can be improved; however, the depth of focus and thus the imaging range will be sacrificed correspondingly. In this work, we report our development of blind-deconvolution optical-resolution photoacoustic microscopy (BD-PAM) that can provide a lateral resolution ~2-fold finer than that of conventional OR-PAM (3.04 vs. 5.78μm), without physically increasing the systems numerical aperture. The improvement achieved with BD-PAM is demonstrated by imaging graphene nanoparticles and the microvasculature of mice ears in vivo. Our results suggest that BD-PAM may become a valuable tool for many biomedical applications that require both fine spatial resolution and extended depth of focus.

Reflection-mode in vivo photoacoustic microscopy with subwavelength lateral resolution.

We developed a reflection-mode subwavelength-resolution photoacoustic microscopy system capable of imaging optical absorption contrast in vivo. The simultaneous high-resolution and reflection-mode imaging capacity of the system was enabled by delicately configuring a miniature high-frequency ultrasonic transducer tightly under a water-immersion objective with numerical aperture of 1.0. At 532-nm laser illumination, the lateral resolution of the system was measured to be ~320 nm. With this system, subcellular structures of red blood cells and B16 melanoma cells were resolved ex vivo; microvessels, including individual capillaries, in a mouse ear were clearly imaged label-freely in vivo, using the intrinsic optical absorption from hemoglobin. The current study suggests that, the optical-absorption contrast, subwavelength resolution, and reflection-mode ability of the developed photoacoustic microscopy may empower a wide range of biomedical studies for visualizing cellular and/or subcellular structures.

Multi-parametric quantitative microvascular imaging with optical-resolution photoacoustic microscopy in vivo.

Many diseases involve either the formation of new blood vessels (e.g., tumor angiogenesis) or the damage of existing ones (e.g., diabetic retinopathy) at the microcirculation level. Optical-resolution photoacoustic microscopy (OR-PAM), capable of imaging microvessels in 3D in vivo down to individual capillaries using endogenous contrast, has the potential to reveal microvascular information critical to the diagnosis and staging of microcirculation-related diseases. In this study, we have developed a dedicated microvascular quantification (MQ) algorithm for OR-PAM to automatically quantify multiple microvascular morphological parameters in parallel, including the vessel diameter distribution, the microvessel density, the vascular tortuosity, and the fractal dimension. The algorithm has been tested on in vivo OR-PAM images of a healthy mouse, demonstrating high accuracy for microvascular segmentation and quantification. The developed MQ algorithm for OR-PAM may greatly facilitate quantitative imaging of tumor angiogenesis and many other microcirculation related diseases in vivo.

Advances in Imaging Techniques and Genetically Encoded Probes for Photoacoustic Imaging.

Photoacoustic (PA) imaging is a rapidly emerging biomedical imaging modality that is capable of visualizing cellular and molecular functions with high detection sensitivity and spatial resolution in deep tissue. Great efforts and progress have been made on the development of various PA imaging technologies with improved resolution and sensitivity over the past two decades. Various PA probes with high contrast have also been extensively developed, with many important biomedical applications. In comparison with chemical dyes and nanoparticles, genetically encoded probes offer easier labeling of defined cells within tissues or proteins of interest within a cell, have higher stability in vivo, and eliminate the need for delivery of exogenous substances. Genetically encoded probes have thus attracted increasing attention from researchers in engineering and biomedicine. In this review, we aim to provide an overview of the existing PA imaging technologies and genetically encoded PA probes, and describe further improvements in PA imaging techniques and the near-infrared photochromic protein BphP1, the most sensitive genetically encoded probe thus far, as well as the potential biomedical applications of BphP1-based PA imaging in vivo.

Longitudinal label-free optical-resolution photoacoustic microscopy of tumor angiogenesis in vivo

BACKGROUND Optical-resolution photoacoustic microscopy (OR-PAM) is a high-resolution imaging technology capable of label-free imaging of the morphology and functions of the microvasculature in vivo. Previous studies of angiogenesis by OR-PAM were carried out primarily with transgenic mice and the mouse ear model. While important findings have been generated using this approach, the application of OR-PAM to the more widely used subcutaneous dorsal tumor models remains challenging, largely due to the respiratory and cardiac motion artifacts, as well as the protruding tumor contours. METHODS AND MATERIALS A noninvasive dorsal skin-fold (N-DSF) model, along with adaptive z-scanning and a corresponding experimental protocol, is developed. Mammary carcinoma cells (4T1) were administered subcutaneously to the backs of female BALB/c mice for tumor inoculation. The mice were anesthetized using a mixture of isofluorane and oxygen. RESULTS In vivo OR-PAM of angiogenesis with subcutaneous dorsal tumor models in mice has been demonstrated. To test the performance of this method, we have monitored the growth of 4T1 mouse mammary carcinoma in BALB/c mice over a period of 9 days. The major features of tumor angiogenesis, including the change of vascular tortuosity, the dilation of vessel diameters, and the increase of blood supply, have been clearly captured with OR-PAM. CONCLUSIONS In combination with N-DSF model, OR-PAM has demonstrated outstanding capacity to provide label-free monitoring of angiogenesis in tumor. Thus, OR-PAM is of great potential to find broad biomedical applications in the pathophysiological studies of tumor and the treatments for anti-angiogenesis.

Compressed sensing based virtual-detector photoacoustic microscopy in vivo

Abstract. Photoacoustic microscopy (PAM) is becoming a vital tool for various biomedical studies, including functional and molecular imaging of cancer. However, due to the use of a focused ultrasonic transducer for photoacoustic detection, the image quality of conventional PAM degrades rapidly away from the ultrasonic focal zone. To improve the image quality of PAM for out-of-focus regions, we have developed compressed sensing based virtual-detector photoacoustic microscopy (CS-PAM). Through phantom and in vivo experiments, it has been demonstrated that CS-PAM can effectively extend the depth of focus of PAM, and thus may greatly expand its potential biomedical applications.

In vivo photoacoustic/ultrasonic dual-modality endoscopy with a miniaturized full field-of-view catheter

Endoscopy is an essential clinical tool for the diagnosis of gastrointestinal (GI) tract cancer. A photoacoustic system that elegantly combines optical and ultrasound endoscopy advantages by providing high-sensitivity functional information and large imaging depth is a potentially powerful tool for GI tract imaging. Recently, several photoacoustic endoscopic imaging systems have been proposed and developed. However, the relatively large size and rigid length of the catheter make it difficult to translate them into wide clinical applications; while the existing system of a relatively small catheter, capable of in vivo animal imaging, is unable to acquire full (360°) field-of-view cross-section images. In this study, we developed a photoacoustic/ultrasonic dual-modality endoscopic system and a corresponding miniaturized, encapsulated imaging catheter, which provides a full 360° field-of-view. The diameter of the catheter is 2.5 mm, which is compatible with the 2.8-mm instrumental channel of a conventional clinical optical endoscope. Using this system, we demonstrate in vivo 3-dimensional endoscopic photoacoustic/ultrasonic imaging of the colorectum of a healthy Sprague Dawley rat, by depicting vasculature and morphology of the GI tract. The significantly improved imaging field of view, reduced catheter size, high-quality imaging results suggest that the developed photoacoustic/ultrasonic dual-modality endoscopy has a great potential to be translated into a broad range of clinical applications in gastroenterology.

Three-dimensional Hessian matrix-based quantitative vascular imaging of rat iris with optical-resolution photoacoustic microscopy in vivo

Abstract. For the diagnosis and evaluation of ophthalmic diseases, imaging and quantitative characterization of vasculature in the iris are very important. The recently developed photoacoustic imaging, which is ultrasensitive in imaging endogenous hemoglobin molecules, provides a highly efficient label-free method for imaging blood vasculature in the iris. However, the development of advanced vascular quantification algorithms is still needed to enable accurate characterization of the underlying vasculature. We have developed a vascular information quantification algorithm by adopting a three-dimensional (3-D) Hessian matrix and applied for processing iris vasculature images obtained with a custom-built optical-resolution photoacoustic imaging system (OR-PAM). For the first time, we demonstrate in vivo 3-D vascular structures of a rat iris with a the label-free imaging method and also accurately extract quantitative vascular information, such as vessel diameter, vascular density, and vascular tortuosity. Our results indicate that the developed algorithm is capable of quantifying the vasculature in the 3-D photoacoustic images of the iris in-vivo, thus enhancing the diagnostic capability of the OR-PAM system for vascular-related ophthalmic diseases in vivo.