Real-time whole-brain imaging of hemodynamics and oxygenation at micro-vessel resolution with ultrafast wide-field photoacoustic microscopy

High-speed high-resolution imaging of the whole-brain hemodynamics is urgently needed to facilitate the next level of neurovascular research. Image acquisition speed and image quality are crucial to visualizing real-time hemodynamics in complex brain vascular networks, and displaying fast pathophysiological dynamics on a micro and macro-level, enabling advances in current queries in neurovascular and brain metabolism research, including stroke, dementia and acute brain injury. Further, real-time oxygen saturation of hemoglobin (sO2) imaging to differentiate arteries from veins and capture fast-paced oxygen delivery dynamics is needed to solve pertinent questions in these fields and beyond. Here, we present a novel ultrafast functional photoacoustic microscopy (UFF-PAM) to image the whole-brain hemodynamics and oxygen delivery. UFF-PAM takes advantage of several key engineering innovations, including Raman-shifter-based dual-wavelength laser excitation, water-immersible 12-facet-polygon scanner, high-sensitivity ultrasound transducer, and deep-learning-based image upsampling. A volumetric imaging rate of 2 Hz has been achieved over a field of view (FOV) of 11× 7.5 × 1.5 mm3 with a high spatial resolution of ~10 µm. Using the UFF-PAM system, we have demonstrated proof-of-concept functional studies on the mouse brains in response to systemic hypoxia, sodium nitroprusside, and stroke. We observed the mouse brain’s fast morphological and functional changes over the entire cortex, including vasoconstriction, vasodilation, and deoxygenation. More interestingly, for the first time, under the whole-brain FOV and micro-vessel resolution, we captured the vasoconstriction and oxygenation change simultaneously in the spreading depolarization (SD) wave. Our work provides a great potential for fundamental brain research under various pathological and physiological conditions.

Non-invasive and low-artifact in vivo brain imaging by using a scanning acoustic-photoacoustic dual mode microscopy

Photoacoustic imaging is a potential candidate for in-vivo brain imaging, whereas, its imaging performance could be degraded by inhomogeneous multi-layered media, consisted of scalp and skull. In this work, we propose a low-artifact photoacoustic microscopy (LAPAM) scheme, which combines conventional acoustic-resolution photoacoustic microscopy with scanning acoustic microscopy to suppress the reflection artifacts induced by multi-layers. Based on similar propagation characteristics of photoacoustic signals and ultrasonic echoes, the ultrasonic echoes can be employed as the filters to suppress the reflection artifacts to obtain low-artifact photoacoustic images. Phantom experiment is used to validate the effectiveness of this method. Furthermore, LAPAM is applied for in-vivo imaging mouse brain without removing the scalp and the skull. Experimental results show that the proposed method successfully achieves the low-artifact brain image, which demonstrates the practical applicability of LAPAM. This work might improve the photoacoustic imaging quality in many biomedical applications, which involve tissue with complex acoustic properties, such as brain imaging through scalp and skull.

Photoacoustic imaging of 3D-printed vascular networks

Thrombosis in the circulation system can lead to major myocardial infarction and cardiovascular deaths. Understanding thrombosis formation is necessary for developing safe and effective treatments. In this work, using digital light processing (DLP)-based 3D printing, we fabricated sophisticated in vitro models of blood vessels with internal microchannels that can be used for thrombosis studies. In this regard, photoacoustic microscopy (PAM) offers a unique advantage for label-free visualization of the 3D-printed vessel models, with large penetration depth and functional sensitivity. We compared the imaging performances of two PAM implementations: optical-resolution PAM and acoustic-resolution PAM, and investigated 3D printed- vessel structures with different patterns of microchannels. Our results show that PAM can provide clear microchannel structures at depths up to 3.6 mm. We further quantified the blood oxygenation in the 3D-printed vascular models, showing that thrombi had lower oxygenation than the normal blood. We expect that PAM can find broad applications in 3D printing and bioprinting for in vitro studies of various vascular and other diseases.

Toward Real-Time Giga-Voxel Optoacoustic/Photoacoustic Microscopy: GPU-Accelerated Fourier Reconstruction with Quasi-3D Implementation

We propose a GPU-accelerated implementation of frequency-domain synthetic aperture focusing technique (SAFT) employing truncated regularized inverse k-space interpolation. Our implementation achieves sub-1s reconstruction time for data sizes of up to 100 M voxels, providing more than a tenfold decrease in reconstruction time as compared to CPU-based SAFT. We provide an empirical model that can be used to predict the execution time of quasi-3D reconstruction for any data size given the specifications of the computing system.

Ultra-high-speed multi-parametric photoacoustic microscopy

Multi-parametric photoacoustic microscopy (PAM) is uniquely capable of simultaneous, high-resolution mapping of blood hemoglobin concentration, oxygenation, and flow in vivo. However, its speed has been limited by the dense sampling required for blood flow quantification. To overcome this limitation, we have developed an ultra-high-speed multi-parametric PAM system, which enables simultaneous acquisition of ~500 densely sampled B-scans by superposing the rapid laser scanning across the line-shaped focus of a cylindrically focused ultrasonic transducer over the conventional mechanical scan of the optical-acoustic dual foci. A novel optical-acoustic combiner is designed and implemented to accommodate the short working distance of the transducer, enabling convenient confocal alignment of the dual foci in the reflection mode. This new system enables continuous monitoring of microvascular hemoglobin concentration, blood oxygenation, and flow over a 4.5 x 3 mm2 area in the awake mouse brain with high spatial and temporal resolution (6.9 μm and 0.3 Hz, respectively).