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.

/pdf/biomedical-photoacoustic-imaging-4s982wv96j.pdf

Biomedical photoacoustic imaging

Our understanding of the cellular implementation of systems-level neural processes like action, thought and emotion has been limited by the availability of tools to interrogate specific classes of neural cells within intact, living brain tissue. Here we identify and develop an archaeal light-driven chloride pump (NpHR) from Natronomonas pharaonis for temporally precise optical inhibition of neural activity. NpHR allows either knockout of single action potentials, or sustained blockade of spiking. NpHR is compatible with ChR2, the previous optical excitation technology we have described, in that the two opposing probes operate at similar light powers but with well-separated action spectra. NpHR, like ChR2, functions in mammals without exogenous cofactors, and the two probes can be integrated with calcium imaging in mammalian brain tissue for bidirectional optical modulation and readout of neural activity. Likewise, NpHR and ChR2 can be targeted together to Caenorhabditis elegans muscle and cholinergic motor neurons to control locomotion bidirectionally. NpHR and ChR2 form a complete system for multimodal, high-speed, genetically targeted, all-optical interrogation of living neural circuits.

Multimodal fast optical interrogation of neural circuitry

Optical imaging through and inside complex samples is a difficult challenge with important applications in many fields. The fundamental problem is that inhomogeneous samples such as biological tissue randomly scatter and diffuse light, preventing the formation of diffraction-limited images. Despite many recent advances, no current method can perform non-invasive imaging in real-time using diffused light. Here, we show that, owing to the ‘memory-effect’ for speckle correlations, a single high-resolution image of the scattered light, captured with a standard camera, encodes sufficient information to image through visually opaque layers and around corners with diffraction-limited resolution. We experimentally demonstrate single-shot imaging through scattering media and around corners using spatially incoherent light and various samples, from white paint to dynamic biological samples. Our single-shot lensless technique is simple, does not require wavefront-shaping nor time-gated or interferometric detection, and is realized here using a camera-phone. It has the potential to enable imaging in currently inaccessible scenarios. Diffraction-limited imaging in a variety of complex media is realized based on analysis of speckle correlations in light captured using a camera phone.

Non-invasive single-shot imaging through scattering layers and around corners via speckle correlations

Photoacoustic imaging is a promising method for visualizing biological tissues with optical absorbers. This article provides an overview of the rapidly developing field of photoacoustic imaging. Photoacoustics, the physical basis of photoacoustic imaging, is analyzed briefly. The merits of photoacoustic technology, compared with optical imaging and ultrasonic imaging, are described. Various imaging techniques are also discussed, including scanning tomography, computed tomography and original detection of photoacoustic imaging. Finally, some biomedical applications of photoacoustic imaging are summarized.

Photoacoustic imaging in biomedicine

In the field of biomedical optics, optical scattering has traditionally limited the range of imaging within tissue to a depth of one millimetre. A recently developed class of wavefront-shaping techniques now aims to overcome this limit and achieve diffraction-limited control of light beyond one centimetre. By manipulating the spatial profile of an optical field before it enters a scattering medium, it is possible to create a micrometre-scale focal spot deep within tissue. To successfully operate in vivo, these wavefront-shaping techniques typically require feedback from within the biological sample. This Review summarizes recently developed 'guidestar' mechanisms that provide feedback for intra-tissue focusing. Potential applications of guidestar-assisted focusing include optogenetic control over neurons, targeted photodynamic therapy and deep tissue imaging.

/pdf/guidestar-assisted-wavefront-shaping-methods-for-focusing-3pl7c6sufk.pdf

Guidestar-assisted wavefront-shaping methods for focusing light into biological tissue

Photoacoustic imaging is an emerging imaging modality that is based upon the photoacoustic effect. In photoacoustic tomography (PAT), the induced acoustic pressure waves are measured by an array of detectors and used to reconstruct an image of the initial pressure distribution. A common challenge faced in PAT is that the measured acoustic waves can only be sparsely sampled. Reconstructing sparsely sampled data using standard methods results in severe artifacts that obscure information within the image. We propose a modified convolutional neural network (CNN) architecture termed fully dense UNet (FD-UNet) for removing artifacts from two-dimensional PAT images reconstructed from sparse data and compare the proposed CNN with the standard UNet in terms of reconstructed image quality.

Fully Dense UNet for 2-D Sparse Photoacoustic Tomography Artifact Removal

Photoacoustic tomography (PAT) is a non-ionizing imaging modality capable of acquiring high contrast and resolution images of optical absorption at depths greater than traditional optical imaging techniques. Practical considerations with instrumentation and geometry limit the number of available acoustic sensors and their “view” of the imaging target, which result in image reconstruction artifacts degrading image quality. Iterative reconstruction methods can be used to reduce artifacts but are computationally expensive. In this work, we propose a novel deep learning approach termed pixel-wise deep learning (Pixel-DL) that first employs pixel-wise interpolation governed by the physics of photoacoustic wave propagation and then uses a convolution neural network to reconstruct an image. Simulated photoacoustic data from synthetic, mouse-brain, lung, and fundus vasculature phantoms were used for training and testing. Results demonstrated that Pixel-DL achieved comparable or better performance to iterative methods and consistently outperformed other CNN-based approaches for correcting artifacts. Pixel-DL is a computationally efficient approach that enables for real-time PAT rendering and improved image reconstruction quality for limited-view and sparse PAT.

/pdf/limited-view-and-sparse-photoacoustic-tomography-for-4167ot1bry.pdf

Limited-View and Sparse Photoacoustic Tomography for Neuroimaging with Deep Learning.

We demonstrate that laser beams can be converged toward a light-absorbing target through optically diffusive media by using photoacoustic-guided interferometric focusing. The convergence of light is achieved by shaping the wavefront of the incident light with a deformable mirror to maximize the photoacoustic signal, which is proportional to the scattered light intensity at the light absorber.

/pdf/photoacoustic-guided-convergence-of-light-through-optically-2oyv0cz922.pdf

Photoacoustic-guided convergence of light through optically diffusive media.

Shock wave therapy (SWT) refers to the use of focused shock waves for treatment of musculoskeletal indications including plantar fascitis and dystrophic mineralization of tendons and joint capsules. Measurements were made of a SWT device that uses a ballistic source. The ballistic source consists of a handpiece within which compressed air (1-4 bar) is used to fire a projectile that strikes a metal applicator placed on the skin. The projectile generates stress waves in the applicator that transmit as pressure waves into tissue. The acoustic fields from two applicators were measured: one applicator was 15 mm in diameter and the surface slightly convex and the second was 12 mm in diameter the surface was concave. Measurements were made in a water tank and both applicators generated a similar pressure pulse consisting of a rectangular positive phase (4 micros duration and up to 8 MPa peak pressure) followed by a predominantly negative tail (duration of 20 micros and peak negative pressure of -6 MPa), with many oscillations. The rise times of the waveforms were around 1 micros and were shown to be too long for the pulses to be considered shock waves. Measurements of the field indicated that region of high pressure was restricted to the near-field (20-40 mm) of the source and was consistent with the Rayleigh distance. The measured acoustic field did not display focusing supported by calculations, which demonstrated that the radius of curvature of the concave surface was too large to effect a focusing gain. Other SWT devices use electrohydraulic, electromagnetic and piezoelectric sources that do result in focused shock waves. This difference in the acoustic fields means there is potentially a significant mechanistic difference between a ballistic source and other SWT devices.

Acoustic Field of a Ballistic Shock Wave Therapy Device

Photoacoustic imaging is an emerging imaging modality that is based upon the photoacoustic effect. In photoacoustic tomography (PAT), the induced acoustic pressure waves are measured by an array of detectors and used to reconstruct an image of the initial pressure distribution. A common challenge faced in PAT is that the measured acoustic waves can only be sparsely sampled. Reconstructing sparsely sampled data using standard methods results in severe artifacts that obscure information within the image. We propose a modified convolutional neural network (CNN) architecture termed Fully Dense UNet (FD-UNet) for removing artifacts from 2D PAT images reconstructed from sparse data and compare the proposed CNN with the standard UNet in terms of reconstructed image quality.

Parag V. Chitnis

Papers

Fully Dense UNet for 2-D Sparse Photoacoustic Tomography Artifact Removal

Limited-View and Sparse Photoacoustic Tomography for Neuroimaging with Deep Learning.

Photoacoustic-guided convergence of light through optically diffusive media.

Acoustic Field of a Ballistic Shock Wave Therapy Device

Fully Dense UNet for 2D Sparse Photoacoustic Tomography Artifact Removal