In vivo photoacoustic and ultrasonic mapping of rat sentinel lymph nodes with a modified commercial ultrasound imaging system

Sentinel lymph node biopsy (SLNB) has become the standard method for axillary staging in breast cancer patients, relying on invasive identification of sentinel lymph nodes (SLNs) following injection of blue dye and radioactive tracers. While SLNB achieves a low false negative rate (5-10%), it is an invasive procedure requiring ionizing radiation. As an alternative to SLNB, ultrasound-guided fine needle aspiration biopsy has been tested clinically. However, ultrasound alone is unable to accurately identify which lymph nodes are sentinel. Therefore, a non-ionizing and noninvasive detection method for accurate SLN mapping is needed. In this study, we successfully imaged methylene blue dye accumulation in vivo in rat axillary lymph nodes using a Phillips iU22 ultrasound imaging system adapted for photoacoustic imaging with an Nd:YAG pumped, tunable dye laser. Photoacoustic images of rat SLNs clearly identify methylene blue dye accumulation within minutes following intradermal dye injection and co-registered photoacoustic/ultrasound images illustrate lymph node position relative to surrounding anatomy. To investigate clinical translation, the imaging depth was extended up to 2.5 cm by adding chicken breast tissue on top of the rat skin surface. These results raise confidence that photoacoustic imaging can be used clinically for accurate, noninvasive SLN mapping

A brief account of nanoparticle contrast agents for photoacoustic imaging

Photoacoustic imaging (PAI) is a hybrid, nonionizing modality offering excellent spatial resolution, deep penetration, and high soft tissue contrast. In PAI, signal is generated based on the absorption of laser-generated optical energy by endogenous tissues or exogenous contrast agents leading to acoustic emissions detected by an ultrasound transducer. Research in this area over the years has shown that PAI has the ability to provide both physiological and molecular imaging, which can be viewed alone or used in a hybrid modality fashion to extend the anatomic and hemodynamic sensitivities of clinical ultrasound. PAI may be performed using inherent contrast afforded by light absorbing molecules such as hemoglobin, myoglobin, and melanin or exogenous small molecule contrast agent such as near infrared dyes and porphyrins. However, this review summarizes the potential of exogenous nanoparticle-based agents for PAI applications including contrast based on gold particles, carbon nanotubes, and encapsulated copper compounds

Three-dimensional photoacoustic imaging with a clinical two-dimensional matrix ultrasound transducer

Photoacoustic tomography provides both structural and functional imaging in vivo based on optical absorption contrast. A novel imaging system that incorporates a two-dimensional matrix ultrasound probe for combined photoacoustic and ultrasonic three-dimensional (3D) volumetric imaging is presented. The system consists of a tunable dye laser pumped by a Nd:YAG laser, a commercial ultrasound imaging system (Philips iU22) with a two-dimensional matrix transducer (Philips X7-2, 2500 elements, 2-7 MHz), and a multichannel data acquisition system which allows us to acquire RF channel data. Compared with alternative 3D techniques, this system is attractive because it can generate co-registered 3D photoacoustic and ultrasound images without mechanical scanning. Moreover, the lateral resolution along the azimuth and elevational directions are measured to be 0.77 ± 0.06 mm and 0.96 ± 0.06 mm, respectively, based on reconstructed photoacoustic images of phantoms containing individual human hairs. Finally, in vivo 3D photoacoustic sentinel lymph node mapping using methylene blue dye in a rat model is demonstrated

Biodegradable Nitrogen-Doped Carbon Nanodots for Non-Invasive Photoacoustic Imaging and Photothermal Therapy

Multifunctional nanoparticles have been widely investigated for biomedical applications, such as imaging, therapy, and drug delivery. Especially, photoactive nanoparticles have received great attention as theranostic agents because of their heat-generating abilities after exposure to laser irradiation. However, photostability and safety issues have been the technical hurdles for further clinical applications. Here, we designed nitrogen (N)-doped carbon nanodots (N-CNDs) that have strong absorption in the near-infrared region, high photostability, and excellent biodegradability. Optimized N-CNDs can be utilized not only as a new photoacoustic (PA) imaging agent but also as a superior photothermal therapy (PTT) agent in vivo because of their strong optical absorption at a specific wavelength. We used N-CNDs to perform in vivo/ex vivo noninvasive PA imaging of sentinel lymph nodes via local delivery and performed PTT for cancer ablation therapy. Finally, biodegradation and renal clearance were confirmed by performing whole-body PA monitoring and a degradation test

Biodegradable Nitrogen-Doped Carbon Nanodots for Non-Invasive Photoacoustic Imaging and Photothermal Therapy

Multifunctional nanoparticles have been widely investigated for biomedical applications, such as imaging, therapy, and drug delivery. Especially, photoactive nanoparticles have received great attention as theranostic agents because of their heat-generating abilities after exposure to laser irradiation. However, photostability and safety issues have been the technical hurdles for further clinical applications. Here, we designed nitrogen (N)-doped carbon nanodots (N-CNDs) that have strong absorption in the near-infrared region, high photostability, and excellent biodegradability. Optimized N-CNDs can be utilized not only as a new photoacoustic (PA) imaging agent but also as a superior photothermal therapy (PTT) agent in vivo because of their strong optical absorption at a specific wavelength. We used N-CNDs to perform in vivo/ex vivo noninvasive PA imaging of sentinel lymph nodes via local delivery and performed PTT for cancer ablation therapy. Finally, biodegradation and renal clearance were confirmed by performing whole-body PA monitoring and a degradation test.11269Ysciescopu

Contrast agents for molecular photoacoustic imaging.

Photoacoustic imaging (PAI) is an emerging tool that bridges the traditional depth limits of ballistic optical imaging and the resolution limits of diffuse optical imaging. Using the acoustic waves generated in response to the absorption of pulsed laser light, it provides noninvasive images of absorbed optical energy density at depths of several centimeters with a resolution of ∼100 μm. This versatile and scalable imaging modality has now shown potential for molecular imaging, which enables visualization of biological processes with systemically introduced contrast agents. Understanding the relative merits of the vast range of contrast agents available, from small-molecule dyes to gold and carbon nanostructures to liposome encapsulations, is a considerable challenge. Here we critically review the physical, chemical and biochemical characteristics of the existing photoacoustic contrast agents, highlighting key applications and present challenges for molecular PAI.This work was supported by CRUK (Career Establishment Award no. C47594/A16267 to J.W. and S.E.B., Core Funding C14303/A17197 to J.W. and S.E.B.), the European Commission (CIG FP7-PEOPLE- 2013-CIG-630729 to J.W. and S.E.B.), the EPSRC-CRUK Cancer Imaging Centre in Cambridge and Manchester (C197/A16465 to J.W. and S.E.B.), King’s College London and University College London Comprehensive Cancer Imaging Centre Cancer Research UK & Engineering and Physical Sciences Research Council, in association with the Medical Research Council and the Department of Health, UK (P.B.), and the European Union (project FAMOS FP7 ICT, contract 317744 to P.B.).This is the author accepted manuscript. The final version is available from Nature Publishing Group via http://dx.doi.org/10.1038/nmeth.392

Contrast Agents for Photoacoustic and Thermoacoustic Imaging: A Review

Photoacoustic imaging (PAI) and thermoacoustic imaging (TAI) are two emerging biomedical imaging techniques that both utilize ultrasonic signals as an information carrier. Unique advantages of PAI and TAI are their abilities to provide high resolution functional information such as hemoglobin and blood oxygenation and tissue dielectric properties relevant to physiology and pathology. These two methods, however, may have a limited detection depth and lack of endogenous contrast. An exogenous contrast agent is often needed to effectively resolve these problems. Such agents are able to greatly enhance the imaging contrast and potentially break through the imaging depth limit. Furthermore, a receptor-targeted contrast agent could trace the molecular and cellular biological processes in tissues. Thus, photoacoustic and thermoacoustic molecular imaging can be outstanding tools for early diagnosis, precise lesion localization, and molecular typing of various diseases. The agents also could be used for therapy in conjugation with drugs or in photothermal therapy, where it functions as an enhancer for the integration of diagnosis and therapy. In this article, we present a detailed review about various exogenous contrast agents for photoacoustic and thermoacoustic molecular imaging. In addition, challenges and future directions of photoacoustic and thermoacoustic molecular imaging in the field of translational medicine are also discussed

Sentinel Lymph Nodes in the Rat: Noninvasive Photoacoustic and US Imaging with a Clinical US System

Purpose: To evaluate in vivo sentinel lymph node (SLN) mapping by using photoacoustic and ultrasonographic (US) imaging with a modified clinical US imaging system. Materials and Methods: Animal protocols were approved by the Animal Studies Committee. Methylene blue dye accumulation in axillary lymph nodes of seven healthy Sprague-Dawley rats was imaged by using a photoacoustic imaging system adapted from a clinical US imaging system. To investigate clinical translation, the imaging depth was extended up to 2.5 cm by adding chicken or turkey breast on top of the rat skin surface. Three-dimensional photoacoustic images were acquired by mechanically scanning the US transducer and light delivery fiber bundle along the elevational direction. Results: Photoacoustic images of rat SLNs clearly help visualization of methylene blue accumulation, whereas coregistered photoacoustic/US images depict lymph node positions relative to surrounding anatomy. Twenty minutes following methylene blue injection, photoacoustic signals from SLN regions increased nearly 33-fold from baseline signals in preinjection images, and mean contrast between SLNs and background tissue was 76.0 ± 23.7 (standard deviation). Methylene blue accumulation in SLNs was confirmed photoacoustically by using the optical absorption spectrum of the dye. Three-dimensional photoacoustic images demonstrate dynamic accumulation of methylene blue in SLNs after traveling through lymph vessels. Conclusion: In vivo photoacoustic and US mapping of SLNs was successfully demonstrated with a modified clinical US scanner. These results raise confidence that photoacoustic and US imaging can be used clinically for accurate, noninvasive imaging of SLNs for axillary lymph node staging in breast cancer patients

Photoacoustic imaging using nanoclusters

Advances in novel imaging techniques and molecular probes are now extending the opportunity of visualizing molecular targets of disease. Molecular imaging provides anatomic as well as functional and pathological information to sense the expression of molecular biological events. In general, molecular imaging aims to target a specific cell type or tissue and visualize biological events in vivo at the molecular or cellular levels through specific probes. Molecular imaging is usually performed in conjunction with probes for specific targets. The objective of this dissertation is to explore molecular imaging by providing highly efficient photoacoustic nanocluster contrast agents to further validate in vivo molecular imaging, improve the therapeutic procedure, and study fundamental photoacoustic signal processes from cluster of nanoparticles. Initially, a photothermal stimuli-responsive photoacoustic nanocluster was designed and synthesized to provide highly sensitive dynamic contrast within tissue samples. The photoacoustic signal enhancement from clustering of nanoparticles was demonstrated by characterizing the photoacoustic signal from photothermal stimuli-responsive nanoclusters. After characterization, photothermal stimuli-responsive nanoclusters were injected into a mouse tissue and the dynamic photoacoustic response from the nanoclusters activated by an external laser source was observed. This activation can be repeatedly turned on by modulating input laser signals, suggesting a new route for dynamic photoacoustic contrast imaging that will further improve the imaging contrast and more accurately guide the drug release process. Despite tremendous advantages of using these nanoparticles, their safety in a biological environment could be a major hurdle for their in vivo utilization. In order to avoid accumulation and long-term toxicity of nanoparticles, biodegradable nanoclusters consisting of sub-5 nm primary gold particles stabilized by a weakly adsorbed biodegradable polymer were introduced. The photoacoustic signal from biodegradable nanoclusters was quantitatively characterized. In addition, photothermal stability of different sizes of biodegradable nanoclusters was investigated. These nanoclusters were then intravenously injected into mice and biodistribution of nanoparticles was observed. Finally, in vivo spectroscopic photoacoustic imaging was performed on tumor-bearing mice with antibody conjugated biodegradable nanoclusters. This research may provide a new opportunity for molecular imaging to help diagnose tumors at an early stage and promote clinical translation of these techniques.Electrical and Computer Engineerin