Photoacoustic computed tomography (PACT) is a proven technology for imaging hemodynamics in deep brain of small animal models. PACT is inherently compatible with ultrasound (US) imaging, providing complementary contrast mechanisms. While PACT can quantify the brain's oxygen saturation of hemoglobin (sO , US imaging can probe the blood flow based on the Doppler effect. Further, by tracking gas-filled microbubbles, ultrasound localization microscopy (ULM) can map the blood flow velocity with sub-diffraction spatial resolution. In this work, we present a 3D deep-brain imaging system that seamlessly integrates PACT and ULM into a single device, 3D-PAULM. Using a low ultrasound frequency of 4 MHz, 3D-PAULM is capable of imaging the brain hemodynamic functions with intact scalp and skull in a totally non-invasive manner. Using 3D-PAULM, we studied the mouse brain functions with ischemic stroke. Multi-spectral PACT, US B-mode imaging, microbubble-enhanced power Doppler (PD), and ULM were performed on the same mouse brain with intrinsic image co-registration. From the multi-modality measurements, we further quantified blood perfusion, sO2, vessel density, and flow velocity of the mouse brain, showing stroke-induced ischemia, hypoxia, and reduced blood flow. We expect that 3D-PAULM can find broad applications in studying deep brain functions on small animal models.

In photoacoustic computed tomography (PACT) with short-pulsed laser excitation, wideband acoustic signals are generated in biological tissues with frequencies related to the effective shapes and sizes of the optically absorbing targets. Low-frequency photoacoustic signal components correspond to slowly varying spatial features and are often omitted during imaging due to the limited detection bandwidth of the ultrasound transducer, or during image reconstruction as undesired background that degrades image contrast. Here we demonstrate that low-frequency photoacoustic signals, in fact, contain functional and molecular information, and can be used to enhance structural visibility, improve quantitative accuracy, and reduce spare-sampling artifacts. We provide an in-depth theoretical analysis of low-frequency signals in PACT, and experimentally evaluate their impact on several representative PACT applications, such as mapping temperature in photothermal treatment, measuring blood oxygenation in a hypoxia challenge, and detecting photoswitchable molecular probes in deep organs. Our results strongly suggest that low-frequency signals are important for functional and molecular PACT.

Non-invasive small-animal imaging technologies, such as optical imaging, magnetic resonance imaging and x -ray computed tomography, have enabled researchers to study normal biological phenomena or disease progression in their native conditions. However, existing small-animal imaging technologies often lack either the penetration capability for interrogating deep tissues (e.g., optical microscopy), or the functional and molecular sensitivity for tracking specific activities (e.g., magnetic resonance imaging). To achieve functional and molecular imaging in deep tissues, we have developed an integrated photoacoustic, ultrasound and acoustic angiographic tomography (PAUSAT) system by seamlessly combining light and ultrasound. PAUSAT can perform three imaging modes simultaneously with complementary contrast: high-frequency B-mode ultrasound imaging of tissue morphology, microbubble-enabled acoustic angiography of tissue vasculature, and multi-spectral photoacoustic imaging of molecular probes. PAUSAT can provide three-dimensional (3D) multi-contrast images that are co-registered, with high spatial resolutions at large depths. Using PAUSAT, we performed proof-of-concept in vivo experiments on various small animal models: monitoring longitudinal development of placenta and embryo during mouse pregnancy, tracking biodistribution and metabolism of near-infrared organic dye on the whole-body scale, and detecting breast tumor expressing genetically-encoded photoswitchable phytochromes. These results have collectively demonstrated that PAUSAT has broad applicability in biomedical research, providing comprehensive structural, functional, and molecular imaging of small animal models.

Linear-array-based photoacoustic tomography has shown broad applications in biomedical research and preclinical imaging. However, the elevational resolution of a linear array is fundamentally limited due to the weak cylindrical focus of the transducer element. While several methods have been proposed to address this issue, they have all handled the problem in a less time-efficient way. In this work, we propose to improve the elevational resolution of a linear array through Deep-E, a fully dense neural network based on U-net. Deep-E exhibits high computational efficiency by converting the three-dimensional problem into a two-dimension problem: it focused on training a model to enhance the resolution along elevational direction by only using the 2D slices in the axial and elevational plane and thereby reducing the computational burden in simulation and training. We demonstrated the efficacy of Deep-E using various datasets, including simulation, phantom, and human subject results. We found that Deep-E could improve elevational resolution by at least four times and recover the object's true size. We envision that Deep-E will have a significant impact in linear-array-based photoacoustic imaging studies by providing high-speed and high-resolution image enhancement.

One primary technical challenge in photoacoustic microscopy (PAM) is the necessary compromise between spatial resolution and imaging speed. In this study, we propose a novel application of deep learning principles to reconstruct undersampled PAM images and transcend the trade-off between spatial resolution and imaging speed. We compared various convolutional neural network (CNN) architectures, and selected a Fully Dense U-net (FD U-net) model that produced the best results. To mimic various undersampling conditions in practice, we artificially downsampled fully-sampled PAM images of mouse brain vasculature at different ratios. This allowed us to not only definitively establish the ground truth, but also train and test our deep learning model at various imaging conditions. Our results and numerical analysis have collectively demonstrated the robust performance of our model to reconstruct PAM images with as few as 2% of the original pixels, which can effectively shorten the imaging time without substantially sacrificing the image quality.

The penetration depth of photoacoustic imaging in biological tissues has been fundamentally limited by the strong optical attenuation when light is delivered externally through the tissue surface. To address this issue, we previously reported internal-illumination photoacoustic imaging using a customized radial-emission optical fiber diffuser, which, however, has complex fabrication, high cost, and non-uniform light emission. To overcome these shortcomings, we have developed a new type of low-cost fiber diffusers based on a graded-scattering method in which the optical scattering of the fiber diffuser is gradually increased as the light travels. The graded scattering can compensate for the optical attenuation and provide relatively uniform light emission along the diffuser. We performed Monte Carlo numerical simulations to optimize several key design parameters, including the number of scattering segments, scattering anisotropy factor, divergence angle of the optical fiber, and reflective index of the surrounding medium. These optimized parameters collectively result in uniform light emission along the fiber diffuser and can be flexibly adjusted to accommodate different applications. We fabricated and characterized the prototype fiber diffuser made of agarose gel and intralipid. Equipped with the new fiber diffuser, we performed thorough proof-of-concept studies on ex vivo tissue phantoms and an in vivo swine model to demonstrate the deep-imaging capability (~10 cm achieved ex vivo) of photoacoustic tomography. We believe that the internal light delivery via the optimized fiber diffuser is an effective strategy to image deep targets (e.g., kidney) in large animals or humans.

Motor impairment is common following stroke. Diminished strength and coordination contribute to reduced ability to perform activities of daily living. The existing healthcare models focus on delivering rehabilitation during the first few months following stroke. Yet, to regain motor control to the greatest degree, rehabilitation should continue across the lifespan. Currently, individuals with stroke are responsible for self-managing their rehabilitation once therapist guided rehabilitation has concluded. Individuals with stroke are frequently given a written home exercise program to help guide their home rehabilitation, but poor compliance demonstrates a better approach is necessary. In this study, we propose BiGRA, a novel system to more effectively facilitate in-home bilateral rehabilitation. This system holds merits in: (1) An end-to-end task-oriented system for bilateral grip control which emphasizes the modulation of grip coordination between hands; and (2) Innovative metrics framework to quantitatively analyze the motor control performance. The evaluation shows that BiGRA can objectively measure the patients' task performance and is a promising assessment tool for stroke rehabilitation.