Search

Strain elastography and shear wave elastography are commonly used to quantify cervical elasticity. However, the absence of stress information in strain elastography causes difficulty in inter-session elasticity comparison, and the robustness of shear wave elastography is compromised by cervical tissue's high inhomogeneity., Objective: To overcome these limitations, we develop a quantitative cervical elastography system by adding a stress sensor to a clinically used transvaginal ultrasound imaging system., Methods: We record the cervical deformation in B-mode images and measure the probe-surface stress through the sensor. Then we quantify the strain using a customized algorithm and estimate the cervical Young's modulus through stress-strain linear regression., Results: In phantom experiments, we demonstrate the system's high accuracy (alignment with the quasi-static compression method, p-value = 0.369 > 0.05), robustness (alignment between 60°- and 90°-contact measurements, p-value = 0.638 > 0.05), repeatability (consistency of single sonographers' measurements, coefficient of variation < 0.06), and reproducibility (alignment between two sonographers' measurements, Pearson correlation coefficient = 0.981). Applying it to pregnant participants, we observe significant cervical softening (p-value < 0.001): Young's modulus decreases 3.95% weekly and its geometric mean value during the first (11 to 13 weeks), second, and third trimesters are 13.07 kPa, 7.59 kPa, and 4.40 kPa, respectively., Conclusion: The proposed system is accurate, robust, and safe, and enables longitudinal and inter-examiner comparisons., Significance: The system applies to different ultrasound machines with minor software updates, which allows for studies of cervical softening patterns in pregnancy for larger populations, facilitating insights into conditions such as preterm birth.

Transcranial photoacoustic computed tomography presents challenges in human brain imaging due to skull-induced acoustic aberration. Existing full-wave image reconstruction methods rely on a unified elastic wave equation for skull shear and longitudinal wave propagation, therefore demanding substantial computational resources. We propose an efficient discrete imaging model based on finite element discretization. The elastic wave equation for solids is solely applied to the hard-tissue skull region, while the soft-tissue or coupling-medium region that dominates the simulation domain is modeled with the simpler acoustic wave equation for liquids. The solid-liquid interfaces are explicitly modeled with elastic-acoustic coupling. Furthermore, finite element discretization allows coarser, irregular meshes to conform to object geometry. These factors significantly reduce the linear system size by 20 times to facilitate accurate whole-brain simulations with improved speed. We derive a matched forward-adjoint operator pair based on the model to enable integration with various optimization algorithms. We validate the reconstruction framework through numerical simulations and phantom experiments.

The creation of carbonaceous nanoparticles and their dynamics in hydrocarbon flames are still debated in environmental, combustion, and material sciences. In this study, we introduce single-pulse femtosecond laser sheet-compressed ultrafast photography (fsLS-CUP), an ultrafast imaging technique specifically designed to shed light on and capture ultrafast dynamics stemming from interactions between femtosecond lasers and nanoparticles in flames in a single-shot. fsLS-CUP enables the first-time real-time billion frames-per-second (Gfps) simultaneous two-dimensional (2D) imaging of laser-induced fluorescence (LIF) and laser-induced heating (LIH) that are originated from polycyclic aromatic hydrocarbons (PAHs) and soot particles, respectively. Furthermore, fsLS-CUP provides the real-time spatiotemporal map of femtosecond laser-soot interaction as elastic light scattering (ELS) at an astonishing 250 Gfps. In contrast to existing single-shot ultrafast imaging approaches, which are limited to millions of frames per second only and require multiple laser pulses, our method employs only a single pulse and captures the entire dynamics of laser-induced signals at hundreds of Gfps. Using a single pulse does not change the optical properties of nanoparticles for a following pulse, thus allowing reliable spatiotemporal mapping. Moreover, we found that particle inception and growth are derived from precursors. In essence, as an imaging modality, fsLS-CUP offers ultrafast 2D diagnostics, contributing to the fundamental understanding of nanoparticle's inception and broader applications across different fields, such as material science and biomedical engineering., (© 2024. The Author(s).)

Ultrasonography is a vital component of modern clinical care, with handheld probes routinely used for a variety of applications. However, handheld ultrasound imaging is limited by factors such as the partial-body field of view, operator dependency, contact-induced distortion, and lack of transmission contrast. Here, we demonstrate a new system enabling whole-body ultrasound tomography of humans in reflection and transmission modes. To generate 2D isotropically resolved images across the entire cross-section in vivo, we use a custom 512-element circular ultrasound receiver array with a rotating ultrasonic transmitter. We demonstrate this technique in regions such as the abdomen and legs in healthy volunteers. We also showcase two potential clinical extensions. First, we readily observe subcutaneous and preperitoneal abdominal adipose distributions in our images, enabling adipose thickness assessment over the body without ionizing radiation or mechanical deformation. Second, we demonstrate an approach for rapid (seven frame-per-second) biopsy needle localization with respect to internal tissue features. These capabilities make whole-body ultrasound tomography a potential practical tool for clinical needs currently unmet by other modalities., Competing Interests: Competing interests: D.C.G., J.X., G.K., and L.V.W. have a provisional patent application titled “Thermoacoustic and ultrasound tomography”, U.S. Patent App. 18/336,863, through the California Institute of Technology, Pasadena, CA. The rest of the authors declare that they have no competing interests.

Imaging deep haemodynamics non-invasively remains a quest. Although optical imaging techniques can be used to measure blood flow, they are generally limited to imaging within ∼1 mm below the skin's surface. Here we show that such optical diffusion limit can be broken through by leveraging the spatial heterogeneity of blood and its photoacoustic contrast. Specifically, successive single-shot wide-field photoacoustic images of blood vessels can be used to visualize the frame-to-frame propagation of blood and to estimate blood flow speed and direction pixel-wise. The method, which we named photoacoustic vector tomography (PAVT), allows for the quantification of haemodynamics in veins more than 5 mm deep, as we show for regions in the hands and arms of healthy volunteers. PAVT may offer advantages for the diagnosis and monitoring of vascular diseases and for the mapping of the function of the circulatory system., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)

Techniques for imaging haemodynamics use ionizing radiation or contrast agents or are limited by imaging depth (within approximately 1 mm), complex and expensive data-acquisition systems, or low imaging speeds, system complexity or cost. Here we show that ultrafast volumetric photoacoustic imaging of haemodynamics in the human body at up to 1 kHz can be achieved using a single laser pulse and a single element functioning as 6,400 virtual detectors. The technique, which does not require recalibration for different objects or during long-term operation, enables the longitudinal volumetric imaging of haemodynamics in vasculature a few millimetres below the skin's surface. We demonstrate this technique in vessels in the feet of healthy human volunteers by capturing haemodynamic changes in response to vascular occlusion. Single-shot volumetric photoacoustic imaging using a single-element detector may facilitate the early detection and monitoring of peripheral vascular diseases and may be advantageous for use in biometrics and point-of-care testing., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)

Quantum imaging holds potential benefits over classical imaging but has faced challenges such as poor signal-to-noise ratios, low resolvable pixel counts, difficulty in imaging biological organisms, and inability to quantify full birefringence properties. Here, we introduce quantum imaging by coincidence from entanglement (ICE), using spatially and polarization-entangled photon pairs to overcome these challenges. With spatial entanglement, ICE offers higher signal-to-noise ratios, greater resolvable pixel counts, and the ability to image biological organisms. With polarization entanglement, ICE provides quantitative quantum birefringence imaging capability, where both the phase retardation and the principal refractive index axis angle of an object can be remotely and instantly quantified without changing the polarization states of the photons incident on the object. Furthermore, ICE enables 25 times greater suppression of stray light than classical imaging. ICE has the potential to pave the way for quantum imaging in diverse fields, such as life sciences and remote sensing.

Label-free optical imaging employs natural and nondestructive approaches for the visualisation of biomedical samples for both biological assays and clinical diagnosis. Currently, this field revolves around multiple broad technology-oriented communities, each with a specific focus on a particular modality despite the existence of shared challenges and applications. As a result, biologists or clinical researchers who require label-free imaging are often not aware of the most appropriate modality to use. This manuscript presents a comprehensive review of and comparison among different label-free imaging modalities and discusses common challenges and applications. We expect this review to facilitate collaborative interactions between imaging communities, push the field forward and foster technological advancements, biophysical discoveries, as well as clinical detection, diagnosis, and monitoring of disease., Competing Interests: Competing Interests The authors declare no competing interests.

Significance: Over 100 monoclonal antibodies have been approved by the U.S. Food and Drug Administration (FDA) for clinical use; however, a paucity of knowledge exists regarding the injection site behavior of these formulated therapeutics, particularly the effect of antibody, formulation, and tissue at the injection site. A deeper understanding of antibody behavior at the injection site, especially on blood oxygenation through imaging, will help design improved versions of the therapeutics for a wide range of diseases., Aim: The aim of this research is to understand the dynamics of monoclonal antibodies at the injection site as well as how the antibody itself affects the functional characteristics of the injection site [e.g., blood oxygen saturation ( sO 2 )]., Approach: We employed triple-wavelength equipped functional photoacoustic imaging to study the dynamics of dye-labeled and unlabeled monoclonal antibodies at the site of injection in a mouse ear. We injected a near-infrared dye-labeled (and unlabeled) human IgG4 isotype control antibody into the subcutaneous space in mouse ears to analyze the injection site dynamics and quantify molecular movement, as well as its effect on local hemodynamics., Results: We performed pharmacokinetic studies of the antibody in different regions of the mouse body to show that dye labeling does not alter the pharmacokinetic characteristics of the antibody and that mouse ear is a viable model for these initial studies. We explored the movement of the antibody in the interstitial space to show that the bolus area grows by ∼ 300 % over 24 h. We discovered that injection of the antibody transiently reduces the local sO 2 levels in mice after prolonged anesthesia without affecting the total hemoglobin content and oxygen extraction fraction., Conclusions: This finding on local oxygen saturation opens a new avenue of study on the functional effects of monoclonal antibody injections. We also show the suitability of the mouse ear model to study antibody dynamics through high-resolution imaging techniques. We quantified the movement of antibodies at the injection site caused by the interstitial fluid, which could be helpful for designing antibodies with tailored absorption speeds in the future., (© 2023 The Authors.)

A three-dimensional (3D) holographic display (3DHD) can preserve all the volumetric information about an object. However, the poor fidelity of 3DHD constrains its applications. Here, we present an ultrahigh-fidelity 3D holographic display that uses scattering for homogenization of angular spectrum. A scattering medium randomizes the incident photons and homogenizes the angular spectrum distribution. The redistributed field is recorded by a photopolymer film with numerous modulation modes and a half-wavelength scale pixel size. We have experimentally improved the contrast of a focal spot to 6 × 10 6 and tightened its spatial resolution to 0.5 micrometers, respectively ~300 and 4.4 times better than digital approaches. By exploiting the spatial multiplexing ability of the photopolymer and the transmission channel selection capability of the scattering medium, we have realized a dynamic holographic display of 3D spirals consisting of 20 foci across 1 millimeter × 1 millimeter × 26 millimeters with uniform intensity.

The deep ocean, characterized by its immense depths and absence of global positioning system (GPS) functionality, presents considerable challenges for search and rescue missions. Inspired by the geolocalization capabilities of migratory marine animals, Bai et al. present a novel method for underwater geolocalization using the polarization patterns of light in the underwater environment. Emulating a sextant using these underwater polarization patterns, the study determines geolocation in underwater settings. Despite prior skepticism, even in low-visibility waters, these patterns, learned through a deep neural network, provide geolocation accuracies of 55 km at 8 m and 255 km at 50 m. This pioneering approach offers implications for search and rescue and hints at navigation mechanisms in marine animals., (© 2023. Changchun Institute of Optics, Fine Mechanics and Physics (CIOMP), CAS.)

Geometric calibration of ultrasound transducer arrays is critical to optimizing the performance of photoacoustic computed tomography (PACT) systems. We present a geometric calibration method that is applicable to a wide range of PACT systems. We obtain the speed of sound and point source locations using surrogate methods, which results in a linear problem in the transducer coordinates. We characterize the estimation error, which informs our choice of the point source arrangement. We demonstrate our method in a three-dimensional PACT system and show that our method improves the contrast-to-noise ratio, the size, and the spread of point source reconstructions by 80 ± 19 % , 19 ± 3 % , and 7 ± 1 % , respectively. We reconstruct the images of a healthy human breast before and after calibration and find that the calibrated image reveals vasculatures that were previously invisible. Our work introduces a method for geometric calibration in PACT and paves the way for improving PACT image quality., Competing Interests: L.V.W. has a financial interest in Microphotoacoustics, Inc., CalPACT, LLC, and Union Photoacoustic Technologies, Ltd., which, however, did not support this work. The other authors declare no competing interests., (© 2023 The Authors.)

Understanding ultrafast dynamics in the femtosecond timescale plays a pivotal role in fundamental research and technology innovation. Spatiotemporal observation of those events in real-time requires imaging speeds greater than 10 12 frames per second (fps), far beyond the fundamental speed limits of the ubiquitous semiconductor sensor technologies. In addition, a majority of femtosecond events are non-repeatable or difficult-to-repeat since they either work in a highly unstable nonlinear regime or require extreme or rare conditions to initiate. Therefore, the traditional pump-probe imaging approach fails since it heavily depends on precise event repetition. Single-shot ultrafast imaging emerges as the only solution; however, existing techniques cannot reach more than 15×10 12 fps, and they only record an insufficient number of frames. Compressed ultrafast spectral photography (CUSP) is proposed to overcome these limitations. Here, CUSP's full design space is explored by manipulating the ultrashort optical pulse in the active illumination. Via parameter optimization, an extraordinarily fast frame rate of 219×10 12 fps is achieved. This implementation of CUSP is also highly flexible, allowing various combinations of imaging speeds and numbers of frames (several hundred up to 1000) to be readily deployed in diverse scientific studies, such as laser-induced transient birefringence, self-focusing, and filaments in dielectric media., (© 2023 The Authors. Advanced Science published by Wiley-VCH GmbH.)

Entangled biphoton sources exhibit nonclassical characteristics and have been applied to imaging techniques such as ghost imaging, quantum holography, and quantum optical coherence tomography. The development of wide-field quantum imaging to date has been hindered by low spatial resolutions, speeds, and contrast-to-noise ratios (CNRs). Here, we present quantum microscopy by coincidence (QMC) with balanced pathlengths, which enables super-resolution imaging at the Heisenberg limit with substantially higher speeds and CNRs than existing wide-field quantum imaging methods. QMC benefits from a configuration with balanced pathlengths, where a pair of entangled photons traversing symmetric paths with balanced optical pathlengths in two arms behave like a single photon with half the wavelength, leading to a two-fold resolution improvement. Concurrently, QMC resists stray light up to 155 times stronger than classical signals. The low intensity and entanglement features of biphotons in QMC promise nondestructive bioimaging. QMC advances quantum imaging to the microscopic level with significant improvements in speed and CNR toward the bioimaging of cancer cells. We experimentally and theoretically prove that the configuration with balanced pathlengths illuminates an avenue for quantum-enhanced coincidence imaging at the Heisenberg limit., (© 2023. The Author(s).)

Significance: Current photoacoustic (PA) imaging modalities typically require either serial detection with a single-element transducer or parallel detections with an ultrasonic array, indicating a dilemma between system cost and imaging throughput. PA topography through ergodic relay (PATER) was recently developed to address this bottleneck. However, PATER requires object-specific calibration due to varied boundary condition and must be recalibrated through pointwise scanning for each object before measurements, which is time-consuming and severely limits practical application., Aim: We aim to develop a new single-shot PA imaging technique that only requires a one-time calibration for imaging different objects using a single-element transducer., Approach: We develop an imaging method, PA imaging through a spatiotemporal encoder (PAISE), to address the above issue. The spatial information is effectively coded into unique temporal features by the spatiotemporal encoder, which allows for compressive image reconstruction. An ultrasonic waveguide is proposed as a critical element to guide the PA waves from the object into the prism, which effectively accounts for the varied boundary condition of different objects. We further add irregular-shaped edges on the prism to introduce randomized internal reflections and further facilitate the scrambling of acoustic waves., Results: The proposed technique is validated through comprehensive numerical simulations and experiments, and it is demonstrated that PAISE can successfully overcome the changed boundary condition and can image different samples given a single calibration., Conclusions: The proposed PAISE technique is capable of single-shot widefield PA imaging with a single-element transducer and does not require sample-specific calibration, which successfully overcomes the major limitation of previous PATER technology., (© 2023 The Authors.)

Wavefront shaping (WFS) is emerging as a promising tool for controlling and focusing light in complex scattering media. The shaping system's speed, the energy gain of the corrected wavefronts, and the control degrees of freedom (DOF) are the most important metrics for WFS, especially for highly scattering and dynamic samples. Despite recent advances, current methods suffer from trade-offs that limit satisfactory performance to only one or two of these metrics. Here, we report a WFS technique that simultaneously achieves high speed, high energy gain, and high control DOF. By combining photorefractive crystal-based analog optical phase conjugation (AOPC) and stimulated emission light amplification, our technique achieves an energy gain approaching unity, more than three orders of magnitude larger than conventional AOPC. The response time of ~10 μ s with about 10 6 control modes corresponds to an average mode time of about 0.01 ns/mode, which is more than 50 times lower than some of the fastest WFS systems to date. We anticipate that this technique will be instrumental in overcoming the optical diffusion limit in photonics and translate WFS techniques to real-world applications., Competing Interests: Competing Interests L.W. has a financial interest in Microphotoacoustics, Inc., CalPACT, LLC, and Union Photoacoustic Technologies, Ltd., which, however, did not support this work. The other authors declare no conflicts of interest.

Photoacoustic computed tomography (PACT) images optical absorption contrast by detecting ultrasonic waves induced by optical energy deposition in materials such as biological tissues. An ultrasonic transducer array or its scanning equivalent is used to detect ultrasonic waves. The spatial distribution of the transducer elements must satisfy the spatial Nyquist criterion; otherwise, spatial aliasing occurs and causes artifacts in reconstructed images. The spatial Nyquist criterion poses different requirements on the transducer elements' distributions for different locations in the image domain, which has not been studied previously. In this research, we elaborate on the location dependency through spatiotemporal analysis and propose a location-dependent spatiotemporal antialiasing method. By applying this method to PACT in full-ring array geometry, we effectively mitigate aliasing artifacts with minimal effects on image resolution in both numerical simulations and in vivo experiments.

A stable speckle pattern is generated when a coherent beam illuminates a stationary scattering medium that contains numerous scatterers with fixed positions. To date, there has been no valid method to the best of our knowledge for calculating the speckle pattern of a macro medium with a large number of scatterers. Here, a new method based on possible path sampling with corresponding weights and coherent superposition is presented for the simulation of optical field propagation in a scattering medium and output speckle patterns. In this method, a photon is launched onto a medium with fixed scatterers. It propagates in one direction; upon collision with a scatterer, its direction is updated. The procedure is repeated until it exits the medium. A sampled path is obtained in this manner. By repeatedly launching photons, numerous independent optical paths can be sampled. A speckle pattern, corresponding to the probability density of the photon, is formed by the coherent superposition of sufficiently sampled path lengths ending on a receiving screen. This method can be used in sophisticated studies of the influences of medium parameters, motion of scatterers, sample distortions on speckle distributions, and morphological appearances. It can be used for micro-examination of optical fields in scattering media and may inspire new methods and techniques for non-invasive precision detection and diagnosis of scattering media.

Quantum correlation between two parties serves as an important resource in the surging applications of quantum information. The Bell nonlocality and quantum steering have been proposed to describe non-classical correlations against local-hidden-variable and local-hidden-state theories, respectively. To characterize the two types of non-classical correlations, various nonlocality and steering inequalities have been established, and the amount of inequality violation serves as an important indicator for many entanglement-based tasks. Quantum state tomography has been employed for measuring quantum states, while the method requires intensive computation and does not directly verify either nonlocality or steering over the full domain independent of established theories. Here, we experimentally map the full-domain correlation with bipartite states for nonlocality and quantum steering in CHSH scenarios. The measurement of the maps automatically accounts for detection imperfections. Furthermore, we demonstrate the application of the correlation maps in the entanglement-based quantum key distribution protocol with arbitrary bipartite states. The correlation maps show direct measurements and simple interpretations that can answer fundamental questions about nonlocality and quantum steering as well as contribute to quantum information applications in a straightforward manner.

Unburnt hydrocarbon flames produce soot, which is the second biggest contributor to global warming and harmful to human health. The state-of-the-art high-speed imaging techniques, developed to study non-repeatable turbulent flames, are limited to million-frames-per-second imaging rates, falling short in capturing the dynamics of critical species. Unfortunately, these techniques do not provide a complete picture of flame-laser interactions, important for understanding soot formation. Furthermore, thermal effects induced by multiple consecutive pulses modify the optical properties of soot nanoparticles, thus making single-pulse imaging essential. Here, we report single-shot laser-sheet compressed ultrafast photography (LS-CUP) for billion-frames-per-second planar imaging of flame-laser dynamics. We observed laser-induced incandescence, elastic light scattering, and fluorescence of soot precursors - polycyclic aromatic hydrocarbons (PAHs) in real-time using a single nanosecond laser pulse. The spatiotemporal maps of the PAHs emission, soot temperature, primary nanoparticle size, soot aggregate size, and the number of monomers, present strong experimental evidence in support of the theory and modeling of soot inception and growth mechanism in flames. LS-CUP represents a generic and indispensable tool that combines a portfolio of ultrafast combustion diagnostic techniques, covering the entire lifecycle of soot nanoparticles, for probing extremely short-lived (picoseconds to nanoseconds) species in the spatiotemporal domain in non-repeatable turbulent environments. Finally, LS-CUP's unparalleled capability of ultrafast wide-field temperature imaging in real-time is envisioned to unravel mysteries in modern physics such as hot plasma, sonoluminescence, and nuclear fusion., (© 2023. The Author(s).)

Obtaining frozen sections of bone tissue for intraoperative examination is challenging. To identify the bony edge of resection, orthopaedic oncologists therefore rely on pre-operative X-ray computed tomography or magnetic resonance imaging. However, these techniques do not allow for accurate diagnosis or for intraoperative confirmation of the tumour margins, and in bony sarcomas, they can lead to bone margins up to 10-fold wider (1,000-fold volumetrically) than necessary. Here, we show that real-time three-dimensional contour-scanning of tissue via ultraviolet photoacoustic microscopy in reflection mode can be used to intraoperatively evaluate undecalcified and decalcified thick bone specimens, without the need for tissue sectioning. We validate the technique with gold-standard haematoxylin-and-eosin histology images acquired via a traditional optical microscope, and also show that an unsupervised generative adversarial network can virtually stain the ultraviolet-photoacoustic-microscopy images, allowing pathologists to readily identify cancerous features. Label-free and slide-free histology via ultraviolet photoacoustic microscopy may allow for rapid diagnoses of bone-tissue pathologies and aid the intraoperative determination of tumour margins., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)

Complementary to mainstream cardiac imaging modalities for preclinical research, photoacoustic computed tomography (PACT) can provide functional optical contrast with high imaging speed and resolution. However, PACT has not been demonstrated to reveal the dynamics of whole cardiac anatomy or vascular system without surgical procedure (thoracotomy) for tissue penetration. Here, we achieved non-invasive imaging of rat hearts using the recently developed three-dimensional PACT (3D-PACT) platform, demonstrating the regulated illumination and detection schemes to reduce the effects of optical attenuation and acoustic distortion through the chest wall; thereby, enabling unimpeded visualization of the cardiac anatomy and intracardiac hemodynamics following rapidly scanning the heart within 10 s. We further applied 3D-PACT to reveal distinct cardiac structural and functional changes among the healthy, hypertensive, and obese rats, with optical contrast to uncover differences in cardiac chamber size, wall thickness, and hemodynamics. Accordingly, 3D-PACT provides high imaging speed and nonionizing penetration to capture the whole heart for diagnosing the animal models, holding promises for clinical translation to cardiac imaging of human neonates., (© 2023. The Author(s).)

Non-alcoholic fatty liver disease is the most common liver disorder worldwide, which strongly correlates to obesity, diabetes, and metabolic syndromes. Complementary to mainstream liver diagnostic modalities, photoacoustic tomography (PAT) can provide high-speed images with functional optical contrast. However, PAT has not been demonstrated to study fatty liver anatomy with clear volumetric vasculatures. The livers of multiple rats are non-invasively imaged in vivo using the recently developed 3D PAT platform. The system provides isotropically high spatial resolution in 3D space, presenting clear angiographic structures of rat livers without injecting contrast agents. Furthermore, to quantitatively analyze the difference between the livers of lean and obese rats, the authors measured several PAT features and statistical differences between the two groups are observed. In addition to the anatomy, a time-gated strategy is applied to correct respiration-induced motion artifacts and extracted the hemodynamics of major blood vessels during the breathing cycles. This study demonstrates the capabilities of 3D-PAT to reveal both angiographic anatomy and function in rat livers, providing hematogenous information for fatty liver diagnosis. 3D-PAT, as a new tool for preclinical research, warrants further improvements to be transferred to human pediatric liver imaging., (© 2022 The Authors. Advanced Science published by Wiley-VCH GmbH.)

Optical-resolution photoacoustic microscopy (OR-PAM) can visualize wavelength-dependent optical absorption at the cellular level. However, OR-PAM suffers from a limited depth of field (DOF) due to the tight focus of the optical excitation beam, making it challenging to acquire high-resolution images of samples with uneven surfaces or high-quality volumetric images without z-scanning. To overcome this limitation, we propose needle-shaped beam photoacoustic microscopy (NB-PAM), which can extend the DOF to up to ~28-fold Rayleigh lengths via customized diffractive optical elements (DOEs). The DOE generate a needle beam with a well-maintained beam diameter, a uniform axial intensity distribution, and negligible sidelobes. The advantage of using NB-PAM is demonstrated by both histology-like imaging of fresh slide-free organs using a 266 nm laser and in vivo mouse brain vasculature imaging using a 532 nm laser. The approach provides new perspectives for slide-free intraoperative pathological imaging and in-vivo organ-level imaging., Competing Interests: Competing Interests L.V.W. has a financial interest in MicroPhotoAcoustics, CalPACT, and Union Photoacoustic Technologies, which, however, did not support this work. The remaining authors declare no competing financial interests.

Long-duration in vivo simultaneous imaging of multiple anatomical structures is useful for understanding physiological aspects of diseases, informative for molecular optimization in preclinical models, and has potential applications in surgical settings to improve clinical outcomes. Previous studies involving simultaneous imaging of multiple anatomical structures, for example, blood and lymphatic vessels as well as peripheral nerves and sebaceous glands, have used genetically engineered mice, which require expensive and time-consuming methods. Here, an IgG4 isotype control antibody is labeled with a near-infrared dye and injected into a mouse ear to enable simultaneous visualization of blood and lymphatic vessels, peripheral nerves, and sebaceous glands for up to 3 h using photoacoustic microscopy. For multiple anatomical structure imaging, peripheral nerves and sebaceous glands are imaged inside the injected dye-labeled antibody mass while the lymphatic vessels are visualized outside the mass. The efficacy of the contrast agent to label and localize deep medial lymphatic vessels and lymph nodes using photoacoustic computed tomography is demonstrated. The capability of a single injectable contrast agent to image multiple structures for several hours will potentially improve preclinical therapeutic optimization, shorten discovery timelines, and enable clinical treatments., (© 2022 The Authors. Advanced Science published by Wiley-VCH GmbH.)

Many ultrafast phenomena in biology and physics are fundamental to our scientific understanding but have not yet been visualized owing to the extreme speed and sensitivity requirements in imaging modalities. Two examples are the propagation of passive current flows through myelinated axons and electromagnetic pulses through dielectrics, which are both key to information processing in living organisms and electronic devices. Here, we demonstrate differentially enhanced compressed ultrafast photography (Diff-CUP) to directly visualize propagations of passive current flows at approximately 100 m/s along internodes, i.e., continuous myelinated axons between nodes of Ranvier, from Xenopus laevis sciatic nerves and of electromagnetic pulses at approximately 5 × 10 7  m/s through lithium niobate. The spatiotemporal dynamics of both propagation processes are consistent with the results from computational models, demonstrating that Diff-CUP can span these two extreme timescales while maintaining high phase sensitivity. With its ultrahigh speed (picosecond resolution), high sensitivity, and noninvasiveness, Diff-CUP provides a powerful tool for investigating ultrafast biological and physical phenomena., (© 2022. The Author(s).)

Significance: Optical imaging of responses in fluorescently labeled neurons has progressed significantly in recent years. However, there is still a need to monitor neural activities at divergent spatial scales and at depths beyond the optical diffusion limit., Aim: To meet these needs, we aim to develop multiscale photoacoustic tomography (PAT) to image neural activities across spatial scales with a genetically encoded calcium indicator GCaMP., Approach: First, using photoacoustic microscopy, we show that depth-resolved GCaMP signals can be monitored in vivo from a fly brain in response to odor stimulation without depth scanning and even with the cuticle intact. In vivo monitoring of GCaMP signals was also demonstrated in mouse brains. Next, using photoacoustic computed tomography, we imaged neural responses of a mouse brain slice at depths beyond the optical diffusion limit., Results: We provide the first unambiguous demonstration that multiscale PAT can be used to record neural activities in transgenic flies and mice with select neurons expressing GCaMP., Conclusions: Our results indicate that the combination of multiscale PAT and fluorescent neural activity indicators provides a methodology for imaging targeted neurons at various scales.

Recent advances in functional ultrasound imaging (fUS) and photoacoustic tomography (PAT) offer powerful tools for studying brain function. Complementing each other, fUS and PAT, respectively, measure the cerebral blood flow (CBF) and hemoglobin concentrations, allowing synergistic characterization of cerebral hemodynamics. Here, cross-ray ultrasound tomography (CRUST) and its combination with PAT are presented. CRUST employs a virtual point source from a spherically focused ultrasonic transducer (SFUST) to provide widefield excitation at a 4-kHz pulse repetition frequency. A full-ring-shaped ultrasonic transducer array whose imaging plane is orthogonal to the SFUST's acoustic axis receives scattered ultrasonic waves. Superior to conventional fUS, whose sensitivity to blood flow is angle-dependent and low for perpendicular flow, the crossed transmission and panoramic detection fields of CRUST provide omnidirectional sensitivity to CBF. Using CRUST-PAT, the CBF, oxygen saturation, and hemoglobin concentration changes of the mouse brain during sensory stimulation are measured, with a field of view of ≈7 mm in diameter, spatial resolution of ≈170 µm, and temporal resolution of 200 Hz. The results demonstrate CRUST-PAT as a unique tool for studying cerebral hemodynamics., (© 2022 The Authors. Advanced Science published by Wiley-VCH GmbH.)

Background: Worldwide, 10% of babies are born preterm, defined as a live birth before 37 weeks of gestation. Preterm birth is the leading cause of neonatal death, and survivors face lifelong risks of adverse outcomes. New approaches with large sample sizes are needed to identify strategies to predict and prevent preterm birth. The primary aims of the Washington University Prematurity Research Cohort Study were to conduct three prospective projects addressing possible causes of preterm birth and provide data and samples for future research., Study Design: Pregnant patients were recruited into the cohort between January 2017 and January 2020. Consenting patients were enrolled into the study before 20 weeks' gestation and followed through delivery. Participants completed demographic and lifestyle surveys; provided maternal blood, placenta samples, and cord blood; and participated in up to three projects focused on underlying physiology of preterm birth: cervical imaging (Project 1), circadian rhythms (Project 2), and uterine magnetic resonance imaging and electromyometrial imaging (Project 3)., Results: A total of 1260 participants were enrolled and delivered during the study period. Of the participants, 706 (56%) were Black/African American, 494 (39%) were nulliparous, and 185 (15%) had a previous preterm birth. Of the 1260 participants, 1220 (97%) delivered a live infant. Of the 1220 with a live birth, 163 (14.1%) had preterm birth, of which 74 (6.1%) were spontaneous preterm birth. Of the 1220 participants with a live birth, 841 participated in cervical imaging, 1047 contributed data and/or samples on circadian rhythms, and 39 underwent uterine magnetic resonance imaging. Of the 39, 25 underwent electromyometrial imaging., Conclusion: We demonstrate feasibility of recruiting and retaining a diverse cohort in a complex prospective, longitudinal study throughout pregnancy. The extensive clinical, imaging, survey, and biologic data obtained will be used to explore cervical, uterine, and endocrine physiology of preterm birth and can be used to develop novel approaches to predict and prevent preterm birth., Competing Interests: The authors have declared that no competing interests exist.

Optical techniques offer a wide variety of applications as light-matter interactions provide extremely sensitive mechanisms to probe or treat target media. Most of these implementations rely on the usage of ballistic or quasi-ballistic photons to achieve high spatial resolution. However, the inherent scattering nature of light in biological tissues or tissue-like scattering media constitutes a critical obstacle that has restricted the penetration depth of non-scattered photons and hence limited the implementation of most optical techniques for wider applications. In addition, the components of an optical system are usually designed and manufactured for a fixed function or performance. Recent advances in wavefront shaping have demonstrated that scattering- or component-induced phase distortions can be compensated by optimizing the wavefront of the input light pattern through iteration or by conjugating the transmission matrix of the scattering medium. This offers unprecedented opportunities in many applications to achieve controllable optical delivery or detection at depths or dynamically configurable functionalities by using scattering media to substitute conventional optical components. In this article, the recent progress of wavefront shaping in multidisciplinary fields is reviewed, from optical focusing and imaging with scattering media, functionalized devices, modulation of mode coupling, and nonlinearity in multimode fiber to multimode fiber-based applications. Apart from insights into the underlying principles and recent advances in wavefront shaping implementations, practical limitations and roadmap for future development are discussed in depth. Looking back and looking forward, it is believed that wavefront shaping holds a bright future that will open new avenues for noninvasive or minimally invasive optical interactions and arbitrary control inside deep tissues. The high degree of freedom with multiple scattering will also provide unprecedented opportunities to develop novel optical devices based on a single scattering medium (generic or customized) that can outperform traditional optical components., Competing Interests: The authors declare no competing interests., (© 2022 The Author(s).)

Objective: Ultra-rapid insulin formulations control postprandial hyperglycemia; however, inadequate understanding of injection site absorption mechanisms is limiting further advancement. We used photoacoustic imaging to investigate the injection site dynamics of dye-labeled insulin lispro in the Humalog® and Lyumjev® formulations using the murine ear cutaneous model and correlated it with results from unlabeled insulin lispro in pig subcutaneous injection model., Methods: We employed dual-wavelength optical-resolution photoacoustic microscopy to study the absorption and diffusion of the near-infrared dye-labeled insulin lispro in the Humalog and Lyumjev formulations in mouse ears. We mathematically modeled the experimental data to calculate the absorption rate constants and diffusion coefficients. We studied the pharmacokinetics of the unlabeled insulin lispro in both the Humalog and Lyumjev formulations as well as a formulation lacking both the zinc and phenolic preservative in pigs. The association state of insulin lispro in each of the formulations was characterized using SV-AUC and NMR spectroscopy., Results: Through experiments using murine and swine models, we show that the hexamer dissociation rate of insulin lispro is not the absorption rate-limiting step. We demonstrated that the excipients in the Lyumjev formulation produce local tissue expansion and speed both insulin diffusion and microvascular absorption. We also show that the diffusion of insulin lispro at the injection site drives its initial absorption; however, the rate at which the insulin lispro crosses the blood vessels is its overall absorption rate-limiting step., Conclusions: This study provides insights into injection site dynamics of insulin lispro and the impact of formulation excipients. It also demonstrates photoacoustic microscopy as a promising tool for studying protein therapeutics. The results from this study address critical questions around the subcutaneous behavior of insulin lispro and the formulation excipients, which could be useful to make faster and better controlled insulin formulations in the future., (Copyright © 2022 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Clinical oncology can benefit substantially from imaging technologies that reveal physiological characteristics with multiscale observations. Complementing conventional imaging modalities, photoacoustic imaging (PAI) offers rapid imaging (for example, cross-sectional imaging in real time or whole-breast scanning in 10-15 s), scalably high levels of spatial resolution, safe operation and adaptable configurations. Most importantly, this novel imaging modality provides informative optical contrast that reveals details on anatomical, functional, molecular and histological features. In this Review, we describe the current state of development of PAI and the emerging roles of this technology in cancer screening, diagnosis and therapy. We comment on the performance of cutting-edge photoacoustic platforms, and discuss their clinical applications and utility in various clinical studies. Notably, the clinical translation of PAI is accelerating in the areas of macroscopic and mesoscopic imaging for patients with breast or skin cancers, as well as in microscopic imaging for histopathology. We also highlight the potential of future developments in technological capabilities and their clinical implications, which we anticipate will lead to PAI becoming a desirable and widely used imaging modality in oncological research and practice., (© 2022. Springer Nature Limited.)

A superresolution imaging approach that localizes very small targets, such as red blood cells or droplets of injected photoacoustic dye, has significantly improved spatial resolution in various biological and medical imaging modalities. However, this superior spatial resolution is achieved by sacrificing temporal resolution because many raw image frames, each containing the localization target, must be superimposed to form a sufficiently sampled high-density superresolution image. Here, we demonstrate a computational strategy based on deep neural networks (DNNs) to reconstruct high-density superresolution images from far fewer raw image frames. The localization strategy can be applied for both 3D label-free localization optical-resolution photoacoustic microscopy (OR-PAM) and 2D labeled localization photoacoustic computed tomography (PACT). For the former, the required number of raw volumetric frames is reduced from tens to fewer than ten. For the latter, the required number of raw 2D frames is reduced by 12 fold. Therefore, our proposed method has simultaneously improved temporal (via the DNN) and spatial (via the localization method) resolutions in both label-free microscopy and labeled tomography. Deep-learning powered localization PA imaging can potentially provide a practical tool in preclinical and clinical studies requiring fast temporal and fine spatial resolutions., (© 2022. The Author(s).)

Blood-oxygen-level-dependent (BOLD) functional magnetic resonance imaging of the human brain requires bulky equipment for the generation of magnetic fields. Photoacoustic computed tomography obviates the need for magnetic fields by using light and sound to measure deoxyhaemoglobin and oxyhaemoglobin concentrations to then quantify oxygen saturation and blood volumes. Yet, the available imaging speeds, fields of view (FOV), sensitivities and penetration depths have been insufficient for functional imaging of the human brain. Here, we show that massively parallel ultrasonic transducers arranged hemispherically around the human head can produce tomographic images of the brain with a 10-cm-diameter FOV and spatial and temporal resolutions of 350 µm and 2 s, respectively. In patients who had a hemicraniectomy, a comparison of functional photoacoustic computed tomography and 7 T BOLD functional magnetic resonance imaging showed a strong spatial correspondence in the same FOV and a high temporal correlation between BOLD signals and photoacoustic signals, with the latter enabling faster detection of functional activation. Our findings establish the use of photoacoustic computed tomography for human brain imaging., (© 2021. The Author(s), under exclusive licence to Springer Nature Limited.)

Neurophotonics was launched in 2014 coinciding with the launch of the BRAIN Initiative focused on development of technologies for advancement of neuroscience. For the last seven years, Neurophotonics ' agenda has been well aligned with this focus on neurotechnologies featuring new optical methods and tools applicable to brain studies. While the BRAIN Initiative 2.0 is pivoting towards applications of these novel tools in the quest to understand the brain, this status report reviews an extensive and diverse toolkit of novel methods to explore brain function that have emerged from the BRAIN Initiative and related large-scale efforts for measurement and manipulation of brain structure and function. Here, we focus on neurophotonic tools mostly applicable to animal studies. A companion report, scheduled to appear later this year, will cover diffuse optical imaging methods applicable to noninvasive human studies. For each domain, we outline the current state-of-the-art of the respective technologies, identify the areas where innovation is needed, and provide an outlook for the future directions., (© 2022 The Authors.)

Photoacoustic tomography (PAT) with genetically encoded near-infrared probes enables visualization of specific cell populations in vivo at high resolution deeply in biological tissues. However, because of a lack of proper probes, PAT of cellular dynamics remains unexplored. Here, the authors report a near-infrared Forster resonance energy transfer (FRET) biosensor based on a miRFP670-iRFP720 pair of the near-infrared fluorescent proteins, which enables dynamic functional imaging of active biological processes in deep tissues. By photoacoustically detecting the changes in the optical absorption of the miRFP670 FRET-donor, they monitored cell apoptosis in deep tissue at high spatiotemporal resolution using PAT. Specifically, they detected apoptosis in single cells at a resolution of ≈3 µm in a mouse ear tumor, and in deep brain tumors (>3 mm beneath the scalp) of living mice at a spatial resolution of ≈150 µm with a 20 Hz frame rate. These results open the way for high-resolution photoacoustic imaging of dynamic biological processes in deep tissues using NIR biosensors and PAT., (© 2021 The Authors. Advanced Science published by Wiley-VCH GmbH.)

Focusing light into scattering media, although challenging, is highly desirable in many realms. With the invention of time-reversed ultrasonically encoded (TRUE) optical focusing, acousto-optic modulation was demonstrated as a promising guidestar mechanism for achieving noninvasive and addressable optical focusing into scattering media. Here, we report a new ultrasound-assisted technique, ultrasound-induced field perturbation optical focusing, abbreviated as UFP. Unlike in conventional TRUE optical focusing, where only the weak frequency-shifted first-order diffracted photons due to acousto-optic modulation are useful, here UFP leverages the brighter zeroth-order photons diffracted by an ultrasonic guidestar as information carriers to guide optical focusing. We find that the zeroth-order diffracted photons, although not frequency-shifted, do have a field perturbation caused by the existence of the ultrasonic guidestar. By detecting and time-reversing the differential field of the frequency-unshifted photons when the ultrasound is alternately ON and OFF, we can focus light to the position where the field perturbation occurs inside the scattering medium. We demonstrate here that UFP optical focusing has superior performance to conventional TRUE optical focusing, which benefits from the more intense zeroth-order photons. We further show that UFP optical focusing can be easily and flexibly developed into double-shot realization or even single-shot realization, which is desirable for high-speed wavefront shaping. This new method upsets conventional thinking on the utility of an ultrasonic guidestar and broadens the horizon of light control in scattering media. We hope that it provides a more efficient and flexible mechanism for implementing ultrasound-guided wavefront shaping., (© 2021. The Author(s).)

The successes of magnetic resonance imaging and modern optical imaging of human brain function have stimulated the development of complementary modalities that offer molecular specificity, fine spatiotemporal resolution, and sufficient penetration simultaneously. By virtue of its rich optical contrast, acoustic resolution, and imaging depth far beyond the optical transport mean free path (∼1 mm in biological tissues), photoacoustic computed tomography (PACT) offers a promising complementary modality. In this article, PACT for functional human brain imaging is reviewed in its hardware, reconstruction algorithms, in vivo demonstration, and potential roadmap., Competing Interests: L.V.W. has a financial interest in Microphotoacoustics, Inc., CalPACT, LLC, and Union Photoacoustic Technologies, Ltd., which, however, did not support this work. S.N declares no competing interests., (© 2021 Optical Society of America under the terms of the OSA Open Access Publishing Agreement.)

Significance: Acoustically detecting the rich optical absorption contrast in biological tissues, photoacoustic tomography (PAT) seamlessly bridges the functional and molecular sensitivity of optical excitation with the deep penetration and high scalability of ultrasound detection. As a result of continuous technological innovations and commercial development, PAT has been playing an increasingly important role in life sciences and patient care, including functional brain imaging, smart drug delivery, early cancer diagnosis, and interventional therapy guidance., Aim: Built on our 2016 tutorial article that focused on the principles and implementations of PAT, this perspective aims to provide an update on the exciting technical advances in PAT., Approach: This perspective focuses on the recent PAT innovations in volumetric deep-tissue imaging, high-speed wide-field microscopic imaging, high-sensitivity optical ultrasound detection, and machine-learning enhanced image reconstruction and data processing. Representative applications are introduced to demonstrate these enabling technical breakthroughs in biomedical research., Conclusions: We conclude the perspective by discussing the future development of PAT technologies.

Photoacoustic tomography (PAT) has demonstrated versatile biomedical applications, ranging from tracking single cells to monitoring whole-body dynamics of small animals and diagnosing human breast cancer. Currently, PAT has two major implementations: photoacoustic computed tomography (PACT) and photoacoustic microscopy (PAM). PACT uses a multi-element ultrasonic array for parallel detection, which is relatively complex and expensive. In contrast, PAM requires point-by-point scanning with a single-element detector, which has a limited imaging throughput. The trade-off between the system cost and throughput demands a new imaging method. To this end, we have developed photoacoustic topography through an ergodic relay (PATER). PATER can capture a wide-field image with only a single-element ultrasonic detector upon a single laser shot. This protocol describes the detailed procedures for PATER system construction, including component selection, equipment setup and system alignment. A step-by-step guide for in vivo imaging of a mouse brain is provided as an example application. Data acquisition, image reconstruction and troubleshooting procedures are also elaborated. It takes ~130 min to carry out this protocol, including ~60 min for both calibration and snapshot wide-field data acquisition using a laser with a 2-kHz pulse repetition rate. PATER offers low-cost snapshot wide-field imaging of fast dynamics, such as visualizing blood pulse wave propagation and tracking melanoma tumor cell circulation in mice in vivo. We envision that PATER will have wide biomedical applications and anticipate that the compact size of the setup will allow it to be further developed as a wearable device to monitor human vital signs.

Reversibly photoswitchable probes allow for a wide variety of optical imaging applications. In particular, photoswitchable fluorescent probes have significantly facilitated the development of super-resolution microscopy. Recently, stimulated Raman scattering (SRS) imaging, a sensitive and chemical-specific optical microscopy, has proven to be a powerful live-cell imaging strategy. Driven by the advances of newly developed Raman probes, in particular the pre-resonance enhanced narrow-band vibrational probes, electronic pre-resonance SRS (epr-SRS) has achieved super-multiplex imaging with sensitivity down to 250 nM and multiplexity up to 24 colors. However, despite the high demand, photoswitchable Raman probes have yet to be developed. Here, we propose a general strategy for devising photoswitchable epr-SRS probes. Toward this goal, we exploit the molecular electronic and vibrational coupling, in which we switch the electronic states of the molecules to four different states to turn their ground-state epr-SRS signals on and off. First, we showed that inducing transitions to both the electronic excited state and triplet state can effectively diminish the SRS peaks. Second, we revealed that the epr-SRS signals can be effectively switched off in red-absorbing organic molecules through light-facilitated transitions to a reduced state. Third, we identified that photoswitchable proteins with near-infrared photoswitchable absorbance, whose states are modulable with their electronic resonances detunable toward and away from the pump photon energy, can function as the photoswitchable epr-SRS probes with desirable sensitivity (<1 µM) and low photofatigue (>40 cycles). These photophysical characterizations and proof-of-concept demonstrations should advance the development of novel photoswitchable Raman probes and open up the unexplored Raman imaging capabilities.

Over 90% of breast cancer is cured; yet there remain highly aggressive breast cancers that develop rapidly and are extremely difficult to treat, much less prevent. Breast cancers that rapidly develop between breast image screening are called "interval cancers." The efforts of our team focus on identifying multiscale integrated strategies to identify biologically aggressive precancerous breast lesions. Our goal is to identify spatiotemporal changes that occur prior to development of interval breast cancers. To accomplish this requires integration of new technology. Our team has the ability to perform single cell in situ transcriptional profiling, noncontrast biological imaging, mathematical analysis, and nanoscale evaluation of receptor organization and signaling. These technological innovations allow us to start to identify multidimensional spatial and temporal relationships that drive the transition from biologically aggressive precancer to biologically aggressive interval breast cancer. This article is categorized under: Cancer > Computational Models Cancer > Molecular and Cellular Physiology Cancer > Genetics/Genomics/Epigenetics., (© 2020 The Authors. WIREs Systems Biology and Medicine published by Wiley Periodicals LLC.)

One of the primary challenges in breast cancer diagnosis and treatment is intratumor heterogeneity (ITH), i . e ., the coexistence of different genetically and epigenetically distinct malignant cells within the same tumor. Thus, the identification of ITH is critical for designing better treatments and hence to increase patient survival rates. Herein, we report a noninvasive hybrid imaging technology that integrates multitargeted and multiplexed patchy polymeric photoacoustic contrast agents (MTMPPPCAs) with single-impulse panoramic photoacoustic computed tomography (SIP-PACT). The target specificity ability of MTMPPPCAs to distinguish estrogen and progesterone receptor-positive breast tumors was demonstrated through both fluorescence and photoacoustic measurements and validated by tissue pathology analysis. This work provides the proof-of-concept of the MTMPPPCAs/SIP-PACT system to identify ITH in nonmetastatic tumors, with both high molecular specificity and real-time detection capability.

Neoadjuvant chemotherapy (NAC) has contributed to improving breast cancer outcomes, and it would ideally reduce the need for definitive breast surgery in patients who have no residual cancer after NAC treatment. However, there is no reliable noninvasive imaging modality accepted as the routine method to assess response to NAC. Because of the inability to detect complete response, post-NAC surgery remains the standard of care. To overcome this limitation, a single-breath-hold photoacoustic computed tomography (SBH-PACT) system is developed to provide contrast similar to that of contrast-enhanced magnetic resonance imaging, but with much higher spatial and temporal resolution and without injection of contrast chemicals. SBH-PACT images breast cancer patients at three time points: before, during, and after NAC. The analysis of tumor size, blood vascular density, and irregularity in the distribution and morphology of the blood vessels on SBH-PACT accurately identifies response to NAC as confirmed by the histopathological diagnosis. SBH-PACT shows its near-term potential as a diagnostic tool for assessing breast cancer response to systemic treatment by noninvasively measuring the changes in cancer-associated angiogenesis. Further development of SBH-PACT may also enable serial imaging, rather than the use of current invasive biopsies, to diagnose and follow indeterminate breast lesions., Competing Interests: L.V.W. has a financial interest in Microphotoacoustics, Inc., CalPACT, LLC, and Union Photoacoustic Technologies, Ltd., which, however, did not support this work., (© 2021 The Authors. Advanced Science published by Wiley‐VCH GmbH.)