Photoacoustic tomography (PAT) can create multiscale multicontrast images of living biological structures ranging from organelles to organs. This emerging technology overcomes the high degree of scattering of optical photons in biological tissue by making use of the photoacoustic effect. Light absorption by molecules creates a thermally induced pressure jump that launches ultrasonic waves, which are received by acoustic detectors to form images. Different implementations of PAT allow the spatial resolution to be scaled with the desired imaging depth in tissue while a high depth-to-resolution ratio is maintained. As a rule of thumb, the achievable spatial resolution is on the order of 1/200 of the desired imaging depth, which can reach up to 7 centimeters. PAT provides anatomical, functional, metabolic, molecular, and genetic contrasts of vasculature, hemodynamics, oxygen metabolism, biomarkers, and gene expression. We review the state of the art of PAT for both biological and clinical studies and discuss future prospects.

Photoacoustic Tomography: In Vivo Imaging from Organelles to Organs

Photoacoustic (PA) imaging, also called optoacoustic imaging, is a new biomedical imaging modality based on the use of laser-generated ultrasound that has emerged over the last decade. It is a hybrid modality, combining the high-contrast and spectroscopic-based specificity of optical imaging with the high spatial resolution of ultrasound imaging. In essence, a PA image can be regarded as an ultrasound image in which the contrast depends not on the mechanical and elastic properties of the tissue, but its optical properties, specifically optical absorption. As a consequence, it offers greater specificity than conventional ultrasound imaging with the ability to detect haemoglobin, lipids, water and other light-absorbing chomophores, but with greater penetration depth than purely optical imaging modalities that rely on ballistic photons. As well as visualizing anatomical structures such as the microvasculature, it can also provide functional information in the form of blood oxygenation, blood flow and temperature. All of this can be achieved over a wide range of length scales from micrometres to centimetres with scalable spatial resolution. These attributes lend PA imaging to a wide variety of applications in clinical medicine, preclinical research and basic biology for studying cancer, cardiovascular disease, abnormalities of the microcirculation and other conditions. With the emergence of a variety of truly compelling in vivo images obtained by a number of groups around the world in the last 2–3 years, the technique has come of age and the promise of PA imaging is now beginning to be realized. Recent highlights include the demonstration of whole-body small-animal imaging, the first demonstrations of molecular imaging, the introduction of new microscopy modes and the first steps towards clinical breast imaging being taken as well as a myriad of in vivo preclinical imaging studies. In this article, the underlying physical principles of the technique, its practical implementation, and a range of clinical and preclinical applications are reviewed.

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Biomedical photoacoustic imaging

The nonradiative conversion of light energy into heat (photothermal therapy, PTT) or sound energy (photoacoustic imaging, PAI) has been intensively investigated for the treatment and diagnosis of cancer, respectively. By taking advantage of nanocarriers, both imaging and therapeutic functions together with enhanced tumour accumulation have been thoroughly studied to improve the pre-clinical efficiency of PAI and PTT. In this review, we first summarize the development of inorganic and organic nano photothermal transduction agents (PTAs) and strategies for improving the PTT outcomes, including applying appropriate laser dosage, guiding the treatment via imaging techniques, developing PTAs with absorption in the second NIR window, increasing photothermal conversion efficiency (PCE), and also increasing the accumulation of PTAs in tumours. Second, we introduce the advantages of combining PTT with other therapies in cancer treatment. Third, the emerging applications of PAI in cancer-related research are exemplified. Finally, the perspectives and challenges of PTT and PAI for combating cancer, especially regarding their clinical translation, are discussed. We believe that PTT and PAI having noteworthy features would become promising next-generation non-invasive cancer theranostic techniques and improve our ability to combat cancers.

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Photothermal therapy and photoacoustic imaging via nanotheranostics in fighting cancer

Optical microscopy has been a fundamental tool of biological discovery for more than three centuries, but its in vivo tissue imaging ability has been restricted by light scattering to superficial investigations, even when confocal or multiphoton methods are used. Recent advances in optical and optoacoustic (photoacoustic) imaging now allow imaging at depths and resolutions unprecedented for optical methods. These abilities are increasingly important to understand the dynamic interactions of cellular processes at different systems levels, a major challenge of postgenome biology. This Review discusses promising photonic methods that have the ability to visualize cellular and subcellular components in tissues across different penetration scales. The methods are classified into microscopic, mesoscopic and macroscopic approaches, according to the tissue depth at which they operate. Key characteristics associated with different imaging implementations are described and the potential of these technologies in biological applications is discussed.

Going deeper than microscopy: the optical imaging frontier in biology

Photoacoustic tomography (PAT) is probably the fastest-growing area of biomedical imaging technology, owing to its capacity for high-resolution sensing of rich optical contrast in vivo at depths beyond the optical transport mean free path (~1 mm in human skin). Existing high-resolution optical imaging technologies, such as confocal microscopy and two-photon microscopy, have had a fundamental impact on biomedicine but cannot reach the penetration depths of PAT. By utilizing low ultrasonic scattering, PAT indirectly improves tissue transparency up to 1000-fold and consequently enables deeply penetrating functional and molecular imaging at high spatial resolution. Furthermore, PAT promises in vivo imaging at multiple length-scales; it can image subcellular organelles to organs with the same contrast origin — an important application in multiscale systems biology research.

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Multiscale photoacoustic microscopy and computed tomography.

Abstract Photoacoustic remote sensing has been recently developed as an all-optical imaging modality capable of imaging a variety of endogenous contrast agents label-free. Initially predicted laser pulse-induced refractive index perturbation-based interrogation beam reflectivity modulations have been found to be orders of magnitude smaller than those typically observed experimentally. In this report we utilize a 10 million frames-per-second camera to further investigate these predicted reflectivity modulations, while also exploring other potential mechanisms of laser pulse-induced reflectivity modulations. Laser-induced motion is demonstrated both laterally for gold wires suspended and submerged in air and water, respectively, and carbon fibers submerged in water, and axial motion is observed in gold wires submerged in a depth gradient of intralipid solution. This laser-induced sample motion is anticipated to cause reflectivity modulations local to the interrogation beam profile in microscopy set-ups. Non-motion-based maximum intensity modulations of 3% are also observed in gold wires submerged in water, indicating the presence of the originally predicted reflectivity modulations. Overall, these observations are important as they provide a widefield view of laser-pulse interactions unavailable in previous point scanning-based photoacoustic remote sensing microscopy configurations, where observed mechanisms occur on time-scales orders of magnitude faster than equivalent field of view point scanning capabilities. 

https://www.nature.com/articles/s41598-023-30831-5.pdf

Investigating mechanisms of laser pulse-induced reflectivity modulations in photoacoustic remote sensing with a 10 million frames-per-second camera

Recently a realtime photoacoustic microsco py system has been demonstrated. Un fortunately, however, displayed B-scan images were sometimes difficult to interpret as there was little structural context. In this work, we provide structural context for photoacoustic microscopy images by adding ultrasound biomicroscopy as a complementary and synergistic modality. Our system uses a voice-coil translation stage capable of 1 lateral translation, an d can operate in excess of 15 Hz for 1-cm translations, providing up to 30 ultrasound frames per second. The frame-rate of the photoacoustic acquisitions is limited by the 20-Hz pulse-repetition rate of the laser, but can be increased with a faster-repetition-rate laser. Data from the system is streamed in real time from a 2GS/s PCI data acquisition card to the host PC at rates as high as 200 MB/s. The system should prove useful for various in vivo studies, including combined ultrasound Doppler and photoacoustic imaging. Keywords: photoacoustic imaging, high-frequency ultrasound, Q- switched lasers, biophotonics, cardiovascular research

Development of a fast-scanning combined ultrasound-photoacoustic biomicroscope

Two-dimensional arrays enable volumetric ultrasound imaging but have been limited to small aperture size and hence low resolution due to the high cost and complexity of fabrication, addressing, and processing associated with large fully addressed arrays. Here, we propose Costas arrays as a gridded sparse 2-D array architecture for volumetric ultrasound imaging. Costas arrays have exactly one element for every row and column, such that the vector displacement between any pair of elements is unique. These properties ensure aperiodicity, which helps eliminate grating lobes. Compared with previously reported works, we studied the distribution of active elements based on an order-256 Costas layout on a wider aperture ( ${96} \lambda \times {96} \lambda $ at 7.5 MHz center frequency) for high-resolution imaging. Our investigations with focused scanline imaging of point targets and cyst phantoms showed that Costas arrays exhibit lower peak sidelobe levels compared with random sparse arrays of the same size and offer comparable performance in terms of contrast compared with Fermat spiral arrays. In addition, Costas arrays are gridded, which could ease the manufacturing and has one element for each row/column, which enables simple interconnection strategies. Compared with state-of-the-art matrix probes, which are commonly $32\times32$ , the proposed sparse arrays achieve higher lateral resolution and a wider field of view.

Costas Sparse 2-D Arrays for High-Resolution Ultrasound Imaging

Currently, there is an inability to obtain fast realistic label-free virtual histopathological images of tissues. We previously introduced ultraviolet photoacoustic remote sensing microscopy as a method to obtain virtual hematoxylin contrast albeit without the ability to obtain virtual eosin contrast. By utilizing UV scattering as a high-resolution eosin channel we are able to produce complete H&E-like virtual histology of unstained human breast lumpectomy specimen sections. By further leveraging a novel colormap matching algorithm with this UV scattering, we generate H&E-like output that is shown to have strong concordance with true H&E-stained adjacent sections, showing promising diagnostic utility. 

Ultraviolet scattering and photoacoustic remote sensing (sPARS) microscopy for virtual H&E histology

High sensitive detection with lock-in amplification can provide high signal noise ratio even when noise is in orders of magnitude higher than the signal. Here we report to combine lock-in amplification with a novel photoacoustic remote sensing (PARS) technology to achieve high resolution, high contrast, all optical non-contact photoacoustic imaging at depth beyond optical scattering limitation. We demonstrate phantom measurements from PARS with lock-in technology were several orders of magnitude more sensitive than those from PARS with the broadband detection techniques.

Roger J. Zemp

Papers

Investigating mechanisms of laser pulse-induced reflectivity modulations in photoacoustic remote sensing with a 10 million frames-per-second camera

Development of a fast-scanning combined ultrasound-photoacoustic biomicroscope

Costas Sparse 2-D Arrays for High-Resolution Ultrasound Imaging

Ultraviolet scattering and photoacoustic remote sensing (sPARS) microscopy for virtual H&E histology

Photoacoustic remote sensing microscopy with lock-in amplification