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Abnormal alpha‐synuclein (αSyn) and iron accumulation in the brain play an important role in Parkinson's disease (PD). Herein, we aim to visualize αSyn inclusions and iron deposition in the brains of M83 (A53T) mouse models of PD in vivo. The fluorescent pyrimidoindole derivative THK‐565 probe was characterized by means of recombinant fibrils and brains from 10‐ to 11‐month‐old M83 mice. Concurrent wide‐field fluorescence and volumetric multispectral optoacoustic tomography (vMSOT) imaging were subsequently performed in vivo. Structural and susceptibility weighted imaging (SWI) magnetic resonance imaging (MRI) at 9.4 T as well as scanning transmission x‐ray microscopy (STXM) were performed to characterize the iron deposits in the perfused brains. Immunofluorescence and Prussian blue staining were further performed on brain slices to validate the detection of αSyn inclusions and iron deposition. THK‐565 showed increased fluorescence upon binding to recombinant αSyn fibrils and αSyn inclusions in post‐mortem brain slices from patients with PD and M83 mice. Administration of THK‐565 in M83 mice showed higher cerebral retention at 20 and 40 min post‐intravenous injection by wide‐field fluorescence compared to nontransgenic littermate mice, in congruence with the vMSOT findings. SWI/phase images and Prussian blue indicated the accumulation of iron deposits in the brains of M83 mice, presumably in the Fe3+ form, as evinced by the STXM results. In conclusion, we demonstrated in vivo mapping of αSyn by means of noninvasive epifluorescence and vMSOT imaging and validated the results by targeting the THK‐565 label and SWI/STXM identification of iron deposits in M83 mouse brains ex vivo. [ABSTRACT FROM AUTHOR]

Optoacoustic imaging offers label‐free multi‐parametric characterization of cerebrovascular morphology and hemodynamics at depths and spatiotemporal resolution unattainable with optical microscopy. Effective imaging depth can greatly be enhanced by employing photons in the second near‐infrared (NIR‐II) window. However, diminished absorption by hemoglobin along with a lack of suitable contrast agents hinder an efficient application of the technique in this spectral range. Herein, copper sulfide (CuS) micro‐ and nano‐formulations for multi‐scale optoacoustic imaging in the NIR‐II window are introduced. Dynamic contrast enhancement induced by intravenously administered CuS nanoparticles facilitated visualization of blood perfusion in murine cerebrovascular networks. The individual calcium carbonate microparticles carrying CuS are further shown to generate sufficient responses to enable super‐resolution microvascular imaging and blood flow velocity mapping with localization optoacoustic tomography. [ABSTRACT FROM AUTHOR]

Optoacoustic (or photoacoustic) imaging promises micron‐resolution noninvasive bioimaging with much deeper penetration (>cm) than fluorescence. However, optoacoustic imaging of enzyme activity would require loud, photostable, NIR‐absorbing molecular contrast agents, which remain unknown. Most organic molecular contrast agents are repurposed fluorophores, with severe shortcomings of photoinstability or phototoxicity under optoacoustic imaging, as consequences of their slow S1→S0 electronic relaxation. We now report that known fluorophores can be rationally modified to reach ultrafast S1→S0 rates, without much extra molecular complexity, simply by merging them with molecular switches. Here, we merge azobenzene switches with cyanine dyes to give ultrafast relaxation (<10 ps, >100‐fold faster). Without even adapting instrument settings, these azohemicyanines display outstanding improvements in signal longevity (>1000‐fold increase of photostability) and signal loudness (>3‐fold even at time zero). We show why this simple but unexplored design strategy can still offer stronger performance in the future, and can also increase the spatial resolution and the quantitative linearity of photoacoustic response over extended longitudinal imaging. By bringing the world of molecular switches and rotors to bear on problems facing optoacoustic agents, this practical strategy will help to unleash the full potential of optoacoustic imaging in fundamental studies and translational uses. [ABSTRACT FROM AUTHOR]

Optoacoustic (OA) tomography is a powerful noninvasive preclinical imaging tool enabling high resolution whole-body visualization of biodistribution and dynamics of molecular agents. The technique yet lacks endogenous soft-tissue contrast, which often hampers anatomical navigation. Herein, we devise spiral volumetric optoacoustic and ultrasound (SVOPUS) tomography for concurrent OA and pulse-echo ultrasound (US) imaging of whole mice. To this end, a spherical array transducer featuring a central curvilinear segment is employed. Full rotation of the array renders transverse US and OA views, while additional translation facilitates volumetric whole-body imaging with high spatial resolution down to 150 µm and 110 µm in the OA and US modes, respectively. OA imaging revealed blood-filled, vascular organs like heart, liver, spleen, kidneys, and surrounding vasculature, whilst complementary details of bones, lungs, and skin boundaries were provided by the US. The dual-modal capability of SVOPUS for label-free imaging of tissue morphology and function is poised to facilitate pharmacokinetic studies, disease monitoring, and image-guided therapies.

Optoacoustic imaging enables the measurement of tissue oxygen saturation (sO2) and blood perfusion while being utilized for detecting tumor microenvironments. Our aim was to employ multispectral optoacoustic tomography (MSOT) to assess immediate‐early changes of hemoglobin level and sO2 within breast tumors during diverse treatments. Mouse breast cancer models were allocated into four groups: control, everolimus (EVE), paclitaxel (PTX), and photodynamic therapy (PDT). Hemoglobin was quantified daily, as well as sO2 and blood perfusion were verified by immunohistochemical (IHC) staining. MSOT showed a temporal window of enhanced oxygenation and improved perfusion in EVE and PTX groups, while sO2 consistently remained below baseline in PDT. The same results were obtained for the IHC. Therefore, MSOT can monitor tumor hypoxia and indirectly reflect blood perfusion in a non‐invasive and non‐labeled way, which has the potential to monitor breast cancer progression early and enable individualized treatment in clinical practice. [ABSTRACT FROM AUTHOR]

Major biological discoveries are made by interrogating living organisms with light. However, the limited penetration of un‐scattered photons within biological tissues limits the depth range covered by optical methods. Deep‐tissue imaging is achieved by combining light and ultrasound. Optoacoustic imaging exploits the optical generation of ultrasound to render high‐resolution images at depths unattainable with optical microscopy. Recently, laser ultrasound has been suggested as a means of generating broadband acoustic waves for high‐resolution pulse‐echo ultrasound imaging. Herein, an approach is proposed to simultaneously interrogate biological tissues with light and ultrasound based on layer‐by‐layer coating of silica optical fibers with a controlled degree of transparency. The time separation between optoacoustic and ultrasound signals collected with a custom‐made spherical array transducer is exploited for simultaneous 3D optoacoustic and laser ultrasound (OPLUS) imaging with a single laser pulse. OPLUS is shown to enable large‐scale anatomical characterization of tissues along with functional multi‐spectral imaging of chromophores and assessment of cardiac dynamics at ultrafast rates only limited by the pulse repetition frequency of the laser. The suggested approach provides a flexible and scalable means for developing a new generation of systems synergistically combining the powerful capabilities of optoacoustics and ultrasound imaging in biology and medicine. [ABSTRACT FROM AUTHOR]

This work introduces a method of one-dimensional deconvolution with Tikhonov regularization for enhancing three-dimensional optoacoustic images in vivo. The method employs adaptive self-calibration to eliminate frequency-dependent distortions associated with ultrasound propagation and detection. By adapting to the inhomogeneous frequency characteristics of the examined medium, the method eliminates the need for additional calibration experiments. The processing time for three-dimensional optoacoustic data of size 200 × 200 × 100 voxels is less than 5 ms, facilitating the real-time enhancement of angiographic images and improving the effective spatial resolution by more than 50%. [ABSTRACT FROM AUTHOR]

Abstract Major biological discoveries are made by interrogating living organisms with light. However, the limited penetration of un‐scattered photons within biological tissues limits the depth range covered by optical methods. Deep‐tissue imaging is achieved by combining light and ultrasound. Optoacoustic imaging exploits the optical generation of ultrasound to render high‐resolution images at depths unattainable with optical microscopy. Recently, laser ultrasound has been suggested as a means of generating broadband acoustic waves for high‐resolution pulse‐echo ultrasound imaging. Herein, an approach is proposed to simultaneously interrogate biological tissues with light and ultrasound based on layer‐by‐layer coating of silica optical fibers with a controlled degree of transparency. The time separation between optoacoustic and ultrasound signals collected with a custom‐made spherical array transducer is exploited for simultaneous 3D optoacoustic and laser ultrasound (OPLUS) imaging with a single laser pulse. OPLUS is shown to enable large‐scale anatomical characterization of tissues along with functional multi‐spectral imaging of chromophores and assessment of cardiac dynamics at ultrafast rates only limited by the pulse repetition frequency of the laser. The suggested approach provides a flexible and scalable means for developing a new generation of systems synergistically combining the powerful capabilities of optoacoustics and ultrasound imaging in biology and medicine.

The unique optical properties of squaraine dyes make them promising for cancer phototheranostics, but the reported squaraines for in vivo treatments mainly rely on their photothermal effect, where monotherapy cannot achieve the desired therapeutic effect. Here we generated a type of squaraine capable of killing tumors through both photothermal and photodynamic effects. We optimized squaraine structure with selenium modulation and formulated it into nanoparticles that showed strong absorption of infrared light, negligible fluorescence, good photothermal conversion (66.6 %), and strong photodynamic effects even after several irradiation cycles. In addition, the nanoparticles could be tracked through their strong optoacoustic signal. In mice, the nanoparticles effectively accumulated in tumors and eliminated them upon irradiation, without causing adverse effects. Our work demonstrates the potential of selenium modulation of squaraine for precise cancer diagnosis and treatment through synergistic photothermal and photodynamic effects.

Multispectral optoacoustic tomography (MSOT) allows non-invasive molecular disease activity assessment in adults with inflammatory bowel disease (IBD). In this prospective pilot-study, we investigated, whether increased levels of MSOT haemoglobin parameters corresponded to inflammatory activity in paediatric IBD patients, too. 23 children with suspected IBD underwent MSOT of the terminal ileum and sigmoid colon with standard validation (e.g. endoscopy). In Crohn`s disease (CD) and ulcerative colitis (UC) patients with endoscopically confirmed disease activity, MSOT total haemoglobin (HbT) signals were increased in the terminal ileum of CD (72.1 ± 13.0 a.u. vs. 32.9 ± 15.4 a.u., p = 0.0049) and in the sigmoid colon of UC patients (62.9 ± 13.8 a.u. vs. 35.1 ± 16.3 a.u., p = 0.0311) as compared to controls, respectively. Furthermore, MSOT haemoglobin parameters correlated well with standard disease activity assessment (e.g. SES-CD and MSOT HbT (rs =0.69, p = 0.0075). Summarizing, MSOT is a novel technology for non-invasive molecular disease activity assessment in paediatric patients with inflammatory bowel disease.

Objectives: Examination of lymph nodes is one of the most common indications for imaging in the head and neck region. The purpose of this study is to evaluate whether multispectral optoacoustic tomography can be used to observe chromophore differences between benign and malignant neck lymph nodes. Materials and methods: Proof-of-concept ex vivo study of resected cervical lymph nodes from 11 patients. The examination of lymph nodes included imaging with hybrid ultrasound and multispectral tomography system followed by spectral unmixing to separate signals from the endogenous chromophores water, lipid, hemoglobin and oxygenated hemoglobin; calculation of semi-quantitative parameters (total hemoglobin and relative oxygenation of hemoglobin). Comparison of the results from the hybrid measurement with the histopathological results. Results: Most patients suffered from squamous cell carcinoma (n = 7), also metastasis from salivary gland adenocarcinoma and papillary thyroid carcinoma, were included. The comparison between benign cervical lymph nodes and metastases showed significant differences for the absorbers water, lipid, hemoglobin and oxygenated hemoglobin and total hemoglobin. Conclusions: Our ex vivo study suggests that multispectral optoacoustic tomography can be used to detect differences between reactive lymph nodes and metastases. The measurement of endogenous chromophores can be used for this purpose. The examinations are non-invasively and thus potentially improve diagnostic prediction. However, potential influences from the ex vivo setting must be considered. [ABSTRACT FROM AUTHOR]

Multimodal imaging increases the value of stand‐alone modalities by enabling simultaneous multiparametric characterization of biological tissues in vivo. Particularly, optoacoustic (OA) tomography has enabled bringing optical imaging advantages to previously unattainable depths and is currently being hybridized with other imaging technologies for enhanced performance. The full potential of these multimodal imaging approaches can only be achieved with dedicated contrast agents ensuring simultaneous measurements and accurate co‐registration of the information provided. Herein, perfluoropentane‐filled nanodroplets are modified, providing excellent contrast in ultrasound imaging, by introducing shell‐embedded additives offering strong contrast in magnetic resonance and OA imaging. Magnetite nanoparticles and indocyanine green (ICG) dye are simultaneously deposited as a multilayer shell on the droplets with the layer‐by‐layer method. This results in sufficiently high amounts of magnetite (0.54 ± 0.08 mg mL−1) and ICG (0.30 ± 0.08 mg mL−1) for efficient magnetic resonance and OA detection, respectively. The high sensitivity achieved with the developed trimodal nanodroplets is first demonstrated in phantom experiments, after which their potential toxic effects, biodistribution, and clearance are examined in vitro and in vivo. Taken together, the obtained results indicate that the developed multilayer polymer shell nanodroplets are efficient hybrid contrast agents for multimodal biomedical imaging applications. [ABSTRACT FROM AUTHOR]

Understanding the neurobiology of complex behaviors requires measurement of activity in the discrete population of active neurons, neuronal ensembles, which control the behavior. Conventional neuroimaging techniques ineffectively measure neuronal ensemble activity in the brain in vivo because they assess the average regional neuronal activity instead of the specific activity of the neuronal ensemble that mediates the behavior. Our functional molecular photoacoustic tomography (FM-PAT) system allows direct imaging of Fos-dependent neuronal ensemble activation in Fos-LacZ transgenic rats in vivo. We tested four experimental conditions and found increased FM-PAT signal in prefrontal cortical areas in rats undergoing conditioned fear or novel context exposure. A parallel immunofluorescence ex vivo study of Fos expression found similar findings. These findings demonstrate the ability of FM-PAT to measure Fos-expressing neuronal ensembles directly in vivo and support a mechanistic role for the prefrontal cortex in higher-order processing of response to specific stimuli or environmental cues.

With the emergence of sentinel node technology, many patients can be staged histopathologically using lymphatic mapping and selective lymphadenectomy. Structural imaging by using US, CT and MR permits precise measurement of lymph node volume, which is strongly associated with neoplastic involvement. Sentinel lymph node detection has been an ideal field of application for nuclear medicine because anatomical data fails to represent the close connections between the lymphatic system and regional lymph nodes, or, more specifically, to identify the first draining lymph node. Hybrid imaging has demonstrated higher accuracy than standard imaging in SLN visualization on images, but it did not change in terms of surgical detection. New alternatives without ionizing radiations are emerging now from "non-radiological" fields, such as ophthalmology and dermatology, where fluorescence or opto-acoustic imaging, for example, are widely used. In this paper, we will analyze the advantages and limits of the main innovative methods in sentinel lymph node detection, including innovations in lymphoscintigraphy techniques that persist as the gold standard to date. [ABSTRACT FROM AUTHOR]

Various morphological and functional parameters of peripheral nerves and their vascular supply are indicative of pathological changes due to injury or disease. Based on recent improvements in optoacoustic image quality, the ability of multispectral optoacoustic tomography, to investigate the vascular environment and morphology of peripheral nerves is explored in vivo in a pilot study on healthy volunteers in tandem with ultrasound imaging (OPUS). The unique ability of optoacoustic imaging to visualize the vasa nervorum by observing intraneural vessels in healthy nerves is showcased in vivo for the first time. In addition, it is demonstrated that the label‐free spectral optoacoustic contrast of the perfused connective tissue of peripheral nerves can be linked to the endogenous contrast of hemoglobin and collagen. Metrics are introduced to analyze the composition of tissue based on its optoacoustic contrast and show that the high‐resolution spectral contrast reveals specific differences between nervous tissue and reference tissue in the nerve's surrounding. How this showcased extraction of peripheral nerve characteristics using multispectral optoacoustic and ultrasound imaging could offer new insights into the pathophysiology of nerve damage and neuropathies, for example, in the context of diabetes is discussed. [ABSTRACT FROM AUTHOR]

An experimental study on an innovative contrast agent is presented. This work demonstrates that copper sulfide in the form of small-sized nanoparticles can be exploited in photoacoustic imaging. An advantage of this material is strong light absorption in the near-infrared range, especially in the transparency windows of biological tissues. In order to yield a proper contrast, light absorption must be followed by heat release with high efficiency. Therefore, it is important to evaluate the photochemical conversion efficiency of the material. We applied a method that is strictly related to photoacoustic applications. The nanoparticles were produced according to a well-established synthesis. Subsequently, they were diluted in pure water to obtain an extinction <0.2/cm at 1064 nm. The photoacoustic signals, generated by 1064 nm laser excitation, were analyzed as a function of concentration and incident laser energy below 70 μJ /pulse. The signals were carefully compared with those of a reference aqueous solution, containing a light-absorbing ionic solute. Data analysis yielded a light-to-heat conversion efficiency 1.0 (±0.1). We discuss this result by comparison with related studies on other types of copper sulfide nanoparticles, where the conversion efficiency reportedly varied from 33% to 93%. The high value determined in the present study possibly indicates that resonant light scattering and luminescence are negligible for our material system. [ABSTRACT FROM AUTHOR]

Optoacoustic imaging (OAI) enables microscale imaging of endogenous chromophores such as hemoglobin at significantly higher penetration depths compared to other optical imaging technologies. Raster‐scanning optoacoustic mesoscopy (RSOM) has recently been shown to identify superficial microvascular changes associated with human skin pathologies. In animal models, the imaging depth afforded by RSOM can enable entirely new capabilities for noninvasive imaging of vascular structures in the gastrointestinal tract, but exact localization of intra‐abdominal organs is still elusive. Herein the development and application of a novel transrectal absorber guide for RSOM (TAG‐RSOM) is presented to enable accurate transabdominal localization and assessment of colonic vascular networks in vivo. The potential of TAG‐RSOM is demonstrated through application during mild and severe acute colitis in mice. TAG‐RSOM enables visualization of transmural vascular networks, with changes in colon wall thickness, blood volume, and OAI signal intensities corresponding to colitis‐associated inflammatory changes. These findings suggest TAG‐RSOM can provide a novel monitoring tool in preclinical IBD models, refining animal procedures and underlines the capabilities of such technologies to address inflammatory bowel diseases in humans. [ABSTRACT FROM AUTHOR]

A diagnosis of pancreatic cancer carries a 5-y survival rate of less than 10%. Furthermore, the detection of pancreatic cancer occurs most often in later stages of the disease due to its location in the retroperitoneum and lack of symptoms (in most cases) until tumors become more advanced. Once diagnosed, cross-sectional imaging techniques are heavily utilized to determine the tumor stage and the potential for surgical resection. However, a major determinant of resectability is the extent of local vascular involvement of the mesenteric vessels and critical tributaries; current imaging techniques have limited capacity to accurately determine vascular involvement. Surrounding inflammation and fibrosis can be difficult to discriminate from viable tumor, making determination of the degree of vascular involvement unreliable. New innovations in fluorescence and optoacoustic imaging techniques may overcome these limitations and make determination of resectability more accurate. These imaging modalities are able to more clearly discern between viable tumor tissue and non-neoplastic inflammation or desmoplasia, allowing clinicians to more reliably characterize vascular involvement and develop individualized treatment plans for patients. This review will discuss the current imaging techniques used to diagnose pancreatic cancer, the barriers that current techniques raise to accurate staging, and novel fluorescence and optoacoustic imaging techniques that may provide more accurate clinical staging of pancreatic cancer. [ABSTRACT FROM AUTHOR]

Silicon photonics is an emerging platform for acoustic sensing, offering exceptional miniaturization and sensitivity. While efforts have focused on silicon-based resonators, silicon nitride resonators can potentially achieve higher Q-factors, further enhancing sensitivity. In this work, a 30 µm silicon nitride microring resonator was fabricated and coated with an elastomer to optimize acoustic sensitivity and signal fidelity. The resonator was characterized acoustically, and its capability for optoacoustic tomography was demonstrated. An acoustic bandwidth of 120 MHz and a noise-equivalent pressure of ∼ 7 mPa/Hz1/2 were demonstrated. The spatially dependent impulse response agreed with theoretical predictions, and spurious acoustic signals, such as reverberations and surface acoustic waves, had a marginal impact. High image fidelity optoacoustic tomography of a 20 µm knot was achieved, confirming the detector’s imaging capabilities. The results show that silicon nitride offers low signal distortion and high-resolution optoacoustic imaging, proving its versatility for acoustic imaging applications.

Abstract Various morphological and functional parameters of peripheral nerves and their vascular supply are indicative of pathological changes due to injury or disease. Based on recent improvements in optoacoustic image quality, the ability of multispectral optoacoustic tomography, to investigate the vascular environment and morphology of peripheral nerves is explored in vivo in a pilot study on healthy volunteers in tandem with ultrasound imaging (OPUS). The unique ability of optoacoustic imaging to visualize the vasa nervorum by observing intraneural vessels in healthy nerves is showcased in vivo for the first time. In addition, it is demonstrated that the label‐free spectral optoacoustic contrast of the perfused connective tissue of peripheral nerves can be linked to the endogenous contrast of hemoglobin and collagen. Metrics are introduced to analyze the composition of tissue based on its optoacoustic contrast and show that the high‐resolution spectral contrast reveals specific differences between nervous tissue and reference tissue in the nerve's surrounding. How this showcased extraction of peripheral nerve characteristics using multispectral optoacoustic and ultrasound imaging could offer new insights into the pathophysiology of nerve damage and neuropathies, for example, in the context of diabetes is discussed.

Abstract Optoacoustic imaging (OAI) enables microscale imaging of endogenous chromophores such as hemoglobin at significantly higher penetration depths compared to other optical imaging technologies. Raster‐scanning optoacoustic mesoscopy (RSOM) has recently been shown to identify superficial microvascular changes associated with human skin pathologies. In animal models, the imaging depth afforded by RSOM can enable entirely new capabilities for noninvasive imaging of vascular structures in the gastrointestinal tract, but exact localization of intra‐abdominal organs is still elusive. Herein the development and application of a novel transrectal absorber guide for RSOM (TAG‐RSOM) is presented to enable accurate transabdominal localization and assessment of colonic vascular networks in vivo. The potential of TAG‐RSOM is demonstrated through application during mild and severe acute colitis in mice. TAG‐RSOM enables visualization of transmural vascular networks, with changes in colon wall thickness, blood volume, and OAI signal intensities corresponding to colitis‐associated inflammatory changes. These findings suggest TAG‐RSOM can provide a novel monitoring tool in preclinical IBD models, refining animal procedures and underlines the capabilities of such technologies to address inflammatory bowel diseases in humans.

High-intensity focused ultrasound (HIFU) enables localized ablation of biological tissues by capitalizing on the synergistic effects of heating and cavitation. Monitoring of those effects is essential for improving the efficacy and safety of HIFU interventions. Herein, we suggest a hybrid optoacoustic-ultrasound (OPUS) approach for real-time assessment of heating and cavitation processes while providing an essential anatomical reference for accurate localization of the HIFU-induced lesion. Both effects could clearly be observed by exploiting the temperature dependence of optoacoustic (OA) signals and the strong contrast of gas bubbles in pulse-echo ultrasound (US) images. The differences in temperature increase and its rate, as recorded with a thermal camera for different HIFU pressures, evinced the onset of cavitation at the expected pressure threshold. The estimated temperatures based on OA signal variations were also within 10–20 % agreement with the camera readings for temperatures below the coagulation threshold (∼50 °C). Experiments performed in excised tissues as well as in a post-mortem mouse demonstrate that both heating and cavitation effects can be effectively visualized and tracked using the OPUS approach. The good sensitivity of the suggested method for HIFU monitoring purposes was manifested by a significant increase in contrast-to-noise ratio within the ablated region by > 10 dB and > 5 dB for the OA and US images, respectively. The hybrid OPUS-based monitoring approach offers the ease of handheld operation thus can readily be implemented in a bedside setting to benefit several types of HIFU treatments used in the clinics.

Optoacoustic tomography has been established as a powerful modality for preclinical imaging. However, efficient whole-body imaging coverage has not been achieved owing to the arduous requirement for continuous acoustic coupling around the animal. In this work, we introduce panoramic (3600) head-to-tail 3D imaging of mice with spiral volumetric optoacoustic tomography (SVOT). The system combines multi-beam illumination and a dedicated head holder enabling uninterrupted acoustic coupling for whole-body scans. Image fidelity is optimized with self-gated respiratory motion rejection and dual speed-of-sound reconstruction algorithms to attain spatial resolution down to 90 µm. The developed system is thus highly suitable for visualizing rapid biodynamics across scales, such as hemodynamic changes in individual organs, responses to treatments and stimuli, perfusion, total body accumulation, or clearance of molecular agents and drugs with unmatched contrast, spatial and temporal resolution.

Optical spectroscopy and microscopy techniques are widely used for basic studies of living systems. However, their application in clinical practice has two fundamental limitations. First, the depth of probing biological tissues with light is small and varies from tenths to several of millimeters. Secondly, it is difficult to use exogenous labels, which increase the sensitivity and specificity of pathological tissue detection, in vivo measurements on patients. This raises the question of the place of biomedical photonics among other physical diagnostic methods used in clinical practice. This article presents an review of optical methods and relatively new certified commercially available medical devices that use photonics to solve intraoperative diagnostic problems, i.e., the discrimination between pathological and healthy tissue sites in vivo and ex vivo by an endogenous optical response. This work discusses a wide range of medical fields in which researchers and engineers have been able to achieve high rates of sensitivity and specificity in solving the problem of classifying such tissues. The advantages and disadvantages of optical imaging and diagnostic methods, which determine their place in clinical practice, are discussed by the example of intraoperative diagnostics. [ABSTRACT FROM AUTHOR]

Abstract Recent studies suggest that metabolic changes and oxygen deficiency in the central nervous system play an important role in the pathophysiology of multiple sclerosis (MS). In our present study, we investigated the changes in oxygenation and analyzed the vascular perfusion of the spinal cord in a rodent model of MS. We performed multispectral optoacoustic tomography of the lumbar spinal cord before and after an oxygen enhancement challenge in mice with experimental autoimmune encephalomyelitis (EAE), a model for MS. In addition, mice were transcardially perfused with lectin to label the vasculature and their spinal columns were optically cleared, followed by light sheet fluorescence microscopy. To analyze the angioarchitecture of the intact spine, we used VesSAP, a novel deep learning-based framework. In EAE mice, the spinal cord had lower oxygen saturation and hemoglobin concentration compared to healthy mice, indicating compromised perfusion of the spinal cord. Oxygen administration reversed hypoxia in the spinal cord of EAE mice, although the ventral region remained hypoxic. Additionally, despite the increased vascular density, we report a reduction in length and complexity of the perfused vascular network in EAE. Taken together, these findings highlight a new aspect of neuroinflammatory pathology, revealing a significant degree of hypoxia in EAE in vivo that is accompanied by changes in spinal vascular perfusion. The study also introduces optoacoustic imaging as a tractable technique with the potential to further decipher the role of hypoxia in EAE and to monitor it in MS patients.

Approximately 19 % of breast cancer patients undergoing breast conserving surgery (BCS) must return for a secondary surgery due to incomplete tumour removal. Our previous work demonstrated that the lower lipid content, characteristic of tumour tissue, was observed as regions of hypo-intense photoacoustic (PA) contrast. The goal of this work was to evaluate feasibility of a low-frequency, hand-held PA imaging probe for surgical margin assessment based on lipid content differences. Here, we describe (i) the design of a prototype hand-held PA imaging probe, (ii) the effect of limited-bandwidth on image contrast, (iii) accuracy towards hypo-intense contrast detection, (iv) the limited-view characteristics of the single sensor design, and (v) early imaging results of an ex-vivo breast cancer specimen. The probe incorporated a single polyvinylidene fluoride acoustic sensor, a 1-to-4 optical fibre bundle and a polycarbonate axicon lens for light delivery. Imaging results on phantoms designed to mimic positive margins demonstrated the ability to detect gaps in optical absorption as small as 1 mm in width. Compared to images from a near full-view PAI system, the hand-held PAI probe had higher signal to noise ratio but suffered from negativity image artifacts. Lumpectomy specimen imaging showed that strong signals can be obtained from the fatty tissue. Taken together, the results show this imaging approach with a hand-held probe has potential for detection of residual breast cancer tissue during BCS; however, more work is needed to reduce the size of the probe to fit within the surgical cavity.

Photoacoustic tomography (PAT) has the potential to recover morphological and functional tissue properties with high spatial resolution. However, previous attempts to solve the optical inverse problem with supervised machine learning were hampered by the absence of labeled reference data. While this bottleneck has been tackled by simulating training data, the domain gap between real and simulated images remains an unsolved challenge. We propose a novel approach to PAT image synthesis that involves subdividing the challenge of generating plausible simulations into two disjoint problems: (1) Probabilistic generation of realistic tissue morphology, and (2) pixel-wise assignment of corresponding optical and acoustic properties. The former is achieved with Generative Adversarial Networks (GANs) trained on semantically annotated medical imaging data. According to a validation study on a downstream task our approach yields more realistic synthetic images than the traditional model-based approach and could therefore become a fundamental step for deep learning-based quantitative PAT (qPAT).

The immediate diagnosis of severe anemia is crucial for patient outcome. However, reliable non-invasive point-of-care diagnostic tools for e.g., ICU monitoring are currently lacking. Using an advanced Multispectral Optoacoustic Tomography (MSOT) research device, we first substantiated a strong positive correlation of MSOT-signal and absolute hemoglobin concentration ex vivo in blood samples. In a clinical exploratory proof-of-concept study, we then evaluated 19 patients with different severities of anemia and controls by non-invasive in vivo measurement of hemoglobin in the radial artery. Our approach proved excellent in identifying patients with severe anemia triggering RBC transfusion based on a strong positive correlation of MSOT-signal intensity and hemoglobin concentration for 700 nm single wavelength and HbR unmixed MSOT-parameter analysis. In conclusion, our study lays the foundation to further develop MSOT-based real-time quantitative perfusion analyses in follow-up preclinical and clinical imaging studies and as a promising diagnostic tool to improve patient care in the future. DRKS00021442

Detailed insight into the radiation-induced changes in tumor microvasculature is crucial to maximize the efficacy of radiotherapy against breast cancer. Recent advances in imaging have enabled precise targeting of solid lesions. However, intratumoral heterogeneity makes treatment planning and monitoring more challenging. Conventional imaging cannot provide high-resolution observation and longitudinal monitoring of large-scale microvascular in response to radiotherapy directly in deep tissues. Herein, we report on an emerging noninvasive imaging assessment method of morphological and functional tumor microvasculature responses with high spatio-temporal resolution by means of optoacoustic imaging (OAI). In vivo imaging of 4T1 breast tumor response to a conventional fractionated radiotherapy at varying dose (14 × 2 Gy and 3 × 8 Gy) has been performed after 2 weeks following treatment. Remarkably, optoacoustic images can generate richful contrast for the tumor microvascular architecture. Besides, the functional status of tumor microvasculature and tumor oxygenation levels were further estimated using OAI. The results revealed the differential (sizedependent) nature of vascular responses to radiation treatments at varying doses. The vessels exhibited an decrease in their density accompanied by a decline in the number of vascular segments following irradiation, compared to the control group. The measurements further revealed an increase of tumor oxygenation levels for 14 × 2 Gy and 3 × 8 Gy irradiations. Our results suggest that OAI could be used to assess the response to radiotherapy based on changes in the functional andmorphological status of tumormicrovasculature, which are closely linked to the intratumor microenvironment. OAI assessment of the tumor microenvironment such as oxygenation status has the potential to be applied to precise radiotherapy strategy. [ABSTRACT FROM AUTHOR]

Optoacoustic tomography (OAT) has recently been advanced toward ultrafast volumetric imaging frame rates in the kilohertz range. As a result, excessive data processing and storage capacity requirements are increasingly being imposed on the imaging systems. OAT data commonly exhibit significant sparsity across the spatial, temporal or spectral domains, which facilitated the development of compressed sensing algorithms exploiting various sparse acquisition and under-sampling schemes to reduce data rates. However, performance of compressed sensing critically depends on a priori knowledge on the type of acquired data and/or imaged object, commonly resulting in lack of general applicability and unpredictable image quality. In this work, we report on a fast adaptive OAT data compression framework operating on fully sampled tomographic data. It is based on a wavelet packet transform that maximizes the data compression ratio according to the desired signal energy loss. A dedicated reconstruction method was further developed that efficiently renders images directly from the compressed data. Up to 1000x compression ratios were achieved while providing efficient control over the resulting image quality from arbitrary datasets exhibiting diverse spatial, temporal and spectral characteristics. Our approach enables faster and longer acquisitions and facilitates long-term storage of large OAT datasets. [ABSTRACT FROM AUTHOR]

Detecting residual nasopharyngeal carcinoma (rNPC) can be difficult because of the coexistence of occult tumours and post-chemoradiation changes, which poses a challenge for both radiologists and surgeons using current imaging methods. Currently, molecular imaging that precisely targets and visualises particular biomarkers in tumours may exceed the specificity and sensitivity of traditional imaging techniques, providing the potential to distinguish tumours from non-neoplastic lesions. Here, we synthesised a HER2/SR-BI-targeted tracer to efficiently position NPC and guide surgery in living mice. This bispecific tracer contained the following two parts: IRDye 800 CW, as an imaging reagent for both optical and optoacoustic imaging, and a fusion peptide (FY-35), as the targeting reagent. Both in vitro and in vivo tests demonstrated that the tracer had higher accumulation and longer retention (up to 48 h) in tumours than a single-targeted probe, and realised sensitive detection of tumours with a minimum size of 3.9 mm. By visualising the vascular network via a customised handheld optoacoustic scan, our intraoperative fluorescence molecular imaging system provides accurate guidance for intraoperative tumour resection. Integrating the advantages of both optical and optoacoustic scanning in an intraoperative image-guided system, this method holds promise for depicting rNPC and guiding salvage surgery. [ABSTRACT FROM AUTHOR]

Optoacoustic (OA) tomography is a powerful noninvasive preclinical imaging tool enabling high resolution whole-body visualization of biodistribution and dynamics of molecular agents. The technique yet lacks endogenous soft-tissue contrast, which often hampers anatomical navigation. Herein, we devise spiral volumetric optoacoustic and ultrasound (SVOPUS) tomography for concurrent OA and pulse-echo ultrasound (US) imaging of whole mice. To this end, a spherical array transducer featuring a central curvilinear segment is employed. Full rotation of the array renders transverse US and OA views, while additional translation facilitates volumetric whole-body imaging with high spatial resolution down to 150 µm and 110 µm in the OA and US modes, respectively. OA imaging revealed blood-filled, vascular organs like heart, liver, spleen, kidneys, and surrounding vasculature, whilst complementary details of bones, lungs, and skin boundaries were provided by the US. The dual-modal capability of SVOPUS for label-free imaging of tissue morphology and function is poised to facilitate pharmacokinetic studies, disease monitoring, and image-guided therapies., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

The analysis of vascular morphology and functionality enables the assessment of disease activity and therapeutic effects in various pathologies. Raster-scanning optoacoustic mesoscopy (RSOM) is an imaging modality that enables the visualization of superficial vascular networks in vivo. In murine models of colitis, deep vascular networks in the colon wall can be visualized by transrectal absorber guide raster-scanning optoacoustic mesoscopy (TAG-RSOM). In order to accelerate the implementation of this technology in translational studies of inflammatory bowel disease, an image-processing pipeline for TAG-RSOM data has been developed. Using optoacoustic data from a murine model of chemically-induced colitis, different image segmentation methods are compared for visualization and quantification of deep vascular patterns in terms of vascular network length and complexity, blood volume, and vessel diameter. The presented image-processing pipeline for TAG-RSOM enables label-free in vivo assessment of changes in the vascular network in murine colitis with broad applications for inflammatory bowel disease research., (© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.)

Rationale and Objectives: The combination of functional biologic data and imaging appearance has the potential to add diagnostic information to help the radiologist evaluate breast masses in an efficient, effective, and cost-conscious manner. This is the first clinical evaluation of the Gen 2(Model 9100, 8101) Imagio® System to assess image quality with both the stand-alone internal ultrasound (IUS), ultrasound-only transducer, and the Optoacoustic/Ultrasound (OA/US) duplex probe (1,2). This study assesses palpable and non-palpable breast abnormalities in patients who are referred for diagnostic breast ultrasound work-up. This study is intended to confirm the clinical acceptability of modifications made to the Imagio® System ultrasound component following Premarket Approval (PMA) of the Imagio® Gen 1 version., Materials and Methods: This prospective, single-arm, non-randomized study included 38 patients presenting with a palpable lump and/or imaging abnormality detected at a single investigational site. Each patient had the breast, and if indicated, the axillary lymph nodes imaged with the Gen 2 Imagio® system., Results: For patients with SenoGram®-predicted Likelihood of Malignancy (LOM) and pathology available (N = 23), observed sensitivity was 100.0% (9/9) with a confidence interval of (66.4%, 100.0%), using a SenoGram®-predicted False Negative Rate (FNR) cut-off of ≤ 2%. Observed specificity was 64.3% (9/14) (Confidence Interval: 35.1%, 87.2%), using a SenoGram®-predicted FNR cut-off of ≤ 2%. At 98% fixed sensitivity, the specificity (fSp) for OA/US + SG was 100.0% while it was 0.0% for IUS. The absolute gain in fSp was 100.0%., Conclusion: Combining structure with morphology can increase specificity without decreasing sensitivity in a real-world setting., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Kenneth A Kist reports a relationship with Seno Medical Instruments that includes: consulting or advisory. Dr. Pamela Otto is on the Advisory Board for Seno. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Association of University Radiologists. Published by Elsevier Inc. All rights reserved.)

Localization optoacoustic tomography (LOT) has recently emerged as a transformative super-resolution technique breaking through the acoustic diffraction limit in deep-tissue optoacoustic (OA) imaging via individual localization and tracking of particles in the bloodstream. However, strong light absorption in red blood cells has previously restricted per-particle OA detection to relatively large microparticles, ≈5 µm in diameter. Herein, it is demonstrated that submicron-sized porous gold nanoparticles, ≈600 nm in diameter, can be individually detected for noninvasive super-resolution imaging with LOT. Ultra-high-speed bright-field microscopy revealed that these nanoparticles generate microscopic plasmonic vapor bubbles, significantly enhancing opto-acoustic energy conversion through a nano-to-micro size transformation. Comprehensive in vitro and in vivo tests further demonstrated the biocompatibility and biosafety of the particles. By reducing the detectable particle size by an order of magnitude, nanoLOT enables microangiographic imaging with a significantly reduced risk of embolisms from particle aggregation and opens new avenues to visualize how nanoparticles reach vascular and potentially extravascular targets. The performance of nanoLOT for non-invasive imaging of microvascular networks in the murine brain anticipates new insights into neurovascular coupling mechanisms and longitudinal microcirculatory changes associated with neurodegenerative diseases., (© 2024 The Author(s). Small published by Wiley‐VCH GmbH.)

Optoacoustic (OA, photoacoustic) imaging capitalizes on the low scattering of ultrasound within biological tissues to provide optical absorption-based contrast with high resolution at depths not reachable with optical microscopy. For deep tissue imaging applications, OA image formation commonly relies on acoustic inversion of time-resolved tomographic data. The excitation of OA responses and subsequent propagation of ultrasound waves can be mathematically described as a forward model enabling image reconstruction via algebraic inversion. These model-based reconstruction methods have been shown to outperform alternative inversion approaches and can further render OA images from incomplete datasets, strongly distorted signals or other suboptimally recorded data. Herein, we provide a general perspective on model-based OA reconstruction methods, review recent progress, and discuss the performance of the different algorithms under practical imaging scenarios.

The Norway lobster, Nephrops norvegicus, supports a key European fishery. Stock assessments for this species are mostly based on trawling and UnderWater TeleVision (UWTV) surveys. However, N. norvegicus are burrowing organisms and these survey methods are unable to sample or observe individuals in their burrows. To account for this, UWTV surveys generally assume that “1 burrow system = 1 animal”, due to the territorial behavior of N. norvegicus. Nevertheless, this assumption still requires in-situ validation. Here, we outline how to improve the accuracy of current stock assessments for N. norvegicus with novel ecological monitoring technologies, including: robotic fixed and mobile camera-platforms, telemetry, environmental DNA (eDNA), and Artificial Intelligence (AI). First, we outline the present status and threat for overexploitation in N. norvegicus stocks. Then, we discuss how the burrowing behavior of N. norvegicus biases current stock assessment methods. We propose that state-of-the-art stationary and mobile robotic platforms endowed with innovative sensors and complemented with AI tools could be used to count both animals and burrows systems in-situ, as well as to provide key insights into burrowing behavior. Next, we illustrate how multiparametric monitoring can be incorporated into assessments of physiology and burrowing behavior. Finally, we develop a flowchart for the appropriate treatment of multiparametric biological and environmental data required to improve current stock assessment methods.

An experimental study on an innovative contrast agent is presented. This work demonstrates that copper sulfide in the form of small-sized nanoparticles can be exploited in photoacoustic imaging. An advantage of this material is strong light absorption in the near-infrared range, especially in the transparency windows of biological tissues. In order to yield a proper contrast, light absorption must be followed by heat release with high efficiency. Therefore, it is important to evaluate the photochemical conversion efficiency of the material. We applied a method that is strictly related to photoacoustic applications. The nanoparticles were produced according to a well-established synthesis. Subsequently, they were diluted in pure water to obtain an extinction

• Various tailored pig models for Duchenne muscular dystrophy (DMD) were generated. • Porcine DMD models resemble human DMD pathology in an accelerated mode. • Proteomics unraveled molecular derangements in skeletal muscle and myocardium. • Multispectral optoacoustic tomography (MSOT) was validated for detection of muscle fibrosis. • Dystrophin was restored by CRISPR/Cas9-mediated deletion of exon 51 in DMD Δ52 pigs. Duchenne muscular dystrophy (DMD) is a fatal X-linked disease caused by mutations in the DMD gene, leading to complete absence of dystrophin and progressive degeneration of skeletal muscles and heart. Animal models are essential for preclinical evaluation of novel diagnostic procedures and treatment strategies. Gene targeting/editing offers the possibility of developing tailored pig models for monogenic diseases. The first porcine DMD model was generated by deletion of DMD exon 52 (DMD Δ52) in cultured kidney cells, which were used for somatic cell nuclear transfer to produce DMD Δ52 offspring. The animals resembled clinical, biochemical, and pathological hallmarks of DMD, but died before sexual maturity, thus preventing their propagation by breeding. This limitation was overcome by the generation of female heterozygous DMD Δ52 carrier pigs, which allowed the establishment of a large breeding colony. In this overview, we summarize how porcine DMD models have been used for dissecting disease mechanisms, for validating multispectral optoacoustic tomography as an imaging modality for monitoring fibrosis, and for preclinical testing of a CRISPR/Cas9 based approach to restore an intact DMD reading frame. Particular advantages of porcine DMD models include their targeted design and the rapid disease progression with early cardiac involvement, facilitating translational studies in reasonable time frames. [ABSTRACT FROM AUTHOR]

Purpose: Abnormal tau accumulation within the brain plays an important role in tauopathies such as Alzheimer's disease and frontotemporal dementia. High-resolution imaging of tau deposits at the whole-brain scale in animal disease models is highly desired. Methods: We approached this challenge by non-invasively imaging the brains of P301L mice of 4-repeat tau with concurrent volumetric multi-spectral optoacoustic tomography (vMSOT) at ~ 115 μm spatial resolution using the tau-targeted pyridinyl-butadienyl-benzothiazole derivative PBB5 (i.v.). In vitro probe characterization, concurrent vMSOT and epi-fluorescence imaging of in vivo PBB5 targeting (i.v.) was performed in P301L and wild-type mice, followed by ex vivo validation using AT-8 antibody for phosphorylated tau. Results: PBB5 showed specific binding to recombinant K18 tau fibrils by fluorescence assay, to post-mortem Alzheimer's disease brain tissue homogenate by competitive binding against [11C]PBB3 and to tau deposits (AT-8 positive) in post-mortem corticobasal degeneration and progressive supranuclear palsy brains. Dose-dependent optoacoustic and fluorescence signal intensities were observed in the mouse brains following i.v. administration of different concentrations of PBB5. In vivo vMSOT brain imaging of P301L mice showed higher retention of PBB5 in the tau-laden cortex and hippocampus compared to wild-type mice, as confirmed by ex vivo vMSOT, epi-fluorescence, multiphoton microscopy, and immunofluorescence staining. Conclusions: We demonstrated non-invasive whole-brain imaging of tau in P301L mice with vMSOT system using PBB5 at a previously unachieved ~ 115 μm spatial resolution. This platform provides a new tool to study tau spreading and clearance in a tauopathy mouse model, foreseeable in monitoring tau targeting putative therapeutics. [ABSTRACT FROM AUTHOR]

Chronic wounds and amputations are common in chronic kidney disease patients needing hemodialysis (HD). HD is often complicated by drops in blood pressure (BP) called intra-dialytic hypotension. Whether intra-dialytic hypotension is associated with detectable changes in foot perfusion, a risk factor for wound formation and impaired healing remains unknown. Photoacoustic (PA) imaging is ideally suited to study perfusion changes. We scanned the feet of 20 HD and 11 healthy subjects. HD patients were scanned before and after a dialysis session whereas healthy subjects were scanned twice at rest and once after a 10 min exercise period while BP was elevated. Healthy (r = 0.70, p

• Improved detection Capabilities: photoacoustic imaging shows enhanced sensitivity and specificity compared to traditional PSA and biopsy methods for prostate cancer. • Real-Time Guidance: enables precise guidance for prostate cancer interventions like biopsies and brachytherapy. • Enhanced monitoring of Therapies: enables more accurate, non-invasive temperature monitoring to optimize thermal treatments like cryotherapy. • Innovative Developments: incorporates advancements such as transurethral illumination, novel contrast agents, and machine learning to improve depth penetration and data analysis. The global health issue of prostate cancer (PCa) requires better diagnosis and treatment. Photoacoustic imaging (PAI) may change PCa management. This review examines PAI's principles, diagnostic role, and therapeutic guidance. PAI uses optical light excitation and ultrasonic detection for high-resolution functional and molecular imaging. PAI uses endogenous and exogenous contrast agents to distinguish cancerous and benign prostate tissues with greater sensitivity and specificity than PSA testing and TRUS-guided biopsy. In addition to diagnosing, PAI can guide and monitor PCa therapy. Its real-time imaging allows precise biopsies and brachytherapy seed placement. Photoacoustic temperature imaging allows non-invasive monitoring of thermal therapies like cryotherapy, improving treatment precision and success. Transurethral illumination probes, innovative contrast agents, integration with other imaging modalities, and machine learning analysis are being developed to overcome depth and data complexity restrictions. PAI could become an essential tool for PCa diagnosis and therapeutic guidance as the field advances. [ABSTRACT FROM AUTHOR]

Recent studies suggest that metabolic changes and oxygen deficiency in the central nervous system play an important role in the pathophysiology of multiple sclerosis (MS). In our present study, we investigated the changes in oxygenation and analyzed the vascular perfusion of the spinal cord in a rodent model of MS. We performed multispectral optoacoustic tomography of the lumbar spinal cord before and after an oxygen enhancement challenge in mice with experimental autoimmune encephalomyelitis (EAE), a model for MS. In addition, mice were transcardially perfused with lectin to label the vasculature and their spinal columns were optically cleared, followed by light sheet fluorescence microscopy. To analyze the angioarchitecture of the intact spine, we used VesSAP, a novel deep learning-based framework. In EAE mice, the spinal cord had lower oxygen saturation and hemoglobin concentration compared to healthy mice, indicating compromised perfusion of the spinal cord. Oxygen administration reversed hypoxia in the spinal cord of EAE mice, although the ventral region remained hypoxic. Additionally, despite the increased vascular density, we report a reduction in length and complexity of the perfused vascular network in EAE. Taken together, these findings highlight a new aspect of neuroinflammatory pathology, revealing a significant degree of hypoxia in EAE in vivo that is accompanied by changes in spinal vascular perfusion. The study also introduces optoacoustic imaging as a tractable technique with the potential to further decipher the role of hypoxia in EAE and to monitor it in MS patients. [ABSTRACT FROM AUTHOR]

Chronic wounds are a major health problem that cause the medical infrastructure billions of dollars every year. Chronic wounds are often difficult to heal and cause significant discomfort. Although wound specialists have numerous therapeutic modalities at their disposal, tools that could three dimensional‐map wound bed physiology and guide therapy do not exist. Visual cues are the current standard but are limited to surface assessment; clinicians rely on experience to predict response to therapy. Photoacoustic (PA) ultrasound (US) is a non‐invasive, hybrid imaging modality that can solve these major limitations. PA relies on the contrast generated by haemoglobin in blood which allows it to map local angiogenesis, tissue perfusion and oxygen saturation—all critical parameters for wound healing. This work evaluates the use of PA‐US to monitor angiogenesis and stratify patients responding versus not‐responding to therapy. We imaged 19 patients with 22 wounds once a week for at least 3 weeks. Our findings suggest that PA imaging directly visualises angiogenesis. Patients responding to therapy showed clear signs of angiogenesis and an increased rate of PA increase (p = 0.002). These responders had a significant and negative correlation between PA intensity and wound size. Hypertension was correlated to impaired angiogenesis in non‐responsive patients. The rate of PA increase and hence the rate of angiogenesis was able to predict healing times within 30 days from the start of monitoring (power = 88%, alpha = 0.05). This early response detection system could help inform management and treatment strategies while improving outcomes and reducing costs. [ABSTRACT FROM AUTHOR]