Photoacoustic Imaging Probe Research Articles

Transrectal Ultrasound and Photoacoustic Imaging Probe for Diagnosis of Prostate Cancer.

A combined transrectal ultrasound and photoacoustic (TRUS–PA) imaging probe was developed for the clear visualization of morphological changes and microvasculature distribution in the prostate, as this is required for accurate diagnosis and biopsy. The probe consisted of a miniaturized 128-element 7 MHz convex array transducer with 134.5° field-of-view (FOV), a bifurcated optical fiber bundle, and two optical lenses. The design goal was to make the size of the TRUS–PA probe similar to that of general TRUS probes (i.e., about 20 mm), for the convenience of the patients. New flexible printed circuit board (FPCB), acoustic structure, and optical lens were developed to meet the requirement of the probe size, as well as to realize a high-performance TRUS–PA probe. In visual assessment, the PA signals obtained with the optical lens were 2.98 times higher than those without the lens. Moreover, the in vivo experiment with the xenograft BALB/c (Albino, Immunodeficient Inbred Strain) mouse model showed that TRUS–PA probe was able to acquire the entire PA image of the mouse tight behind the porcine intestine about 25 mm depth. From the ex vivo and in vivo experimental results, it can be concluded that the developed TRUS–PA probe is capable of improving PA image quality, even though the TRUS–PA probe has a cross-section size and an FOV comparable to those of general TRUS probes.

A pH-responsive photoacoustic imaging probe for tumor pH imaging in vivo based on polyaniline–bovine serum albumin

Precise pH detection in tumors can guide the design of pH-responsive drugs and theranostic agents to improve treatment efficacy. However, most reported pH-responsive probes are fluorescent probes, for which in vivo application is limited by low probe penetration depth. In this study, a pH-responsive polyaniline–bovine serum albumin (BSA) probe was constructed for precise pH detection in tumors using photoacoustic imaging. The probe can be used to generate high-resolution images of deep biological tissues. The photoacoustic signal of the polyaniline–BSA probe exhibits a clear linear relationship with pH in the range of 5–6.8 both in vitro and in vivo, indicating that the probe is ideal for precise pH detection in most tumors. The polyaniline–BSA probe also exhibits satisfactory biocompatibility, low toxicity, fast response, and good reversibility. This work provides a useful in vivo pH detection probe for developing pH-responsive drugs and theranostic agents.

Design and synthesis of a ratiometric photoacoustic imaging probe activated by selenol for visual monitoring of pathological progression of autoimmune hepatitis.

Photoacoustic (PA) imaging with both the high contrast of optical imaging and the high spatial resolution of ultrasound imaging has been regarded as a robust biomedical imaging technique. Autoimmune hepatitis (AIH) is the second largest liver inflammatory disease after viral hepatitis, but its pathogenesis is not fully understood probably due to the lack of an effective in vivo monitoring approach. In this work, an innovative selenol-activated ratiometric PA imaging probe APSel was developed for visual monitoring of pathological progress of AIH. Selenols including selenocysteine (Sec, the major form of Se-containing species in vivo) have been demonstrated to have an effective antioxidant role in inflammation. The reaction of APSel with selenol results in a blue shift of the PA spectrum peak from 860 nm to 690 nm, which enables the ratiometric PA imaging. The APSel probe displays high sensitivity and selectivity to Sec and other selenols. The APSel probe was then employed for ratiometric PA imaging of selenol in cells, and for monitoring the development of AIH in a murine model by tracking the changes of selenol level. The results revealed that the level of selenol was closely correlated with the development of AIH. The proposed APSel, as the first example of a selenol-responsive PA imaging probe, provides a new tool and approach to study and diagnose AIH diseases.

Alkaline phosphatase-triggered self-assembly of near-infrared nanoparticles for the enhanced photoacoustic imaging of tumors.

In this chapter, we discuss the need for the development of enzyme-activatable probes in the field of tumor-targeted photoacoustic (PA) imaging, then we give a brief description of the innovation of designing alkaline phosphatase (ALP)-activatable probes for PA imaging. After that, we provide detailed protocols for the syntheses and characterizations of a near-infrared photoacoustic imaging probe, 1P, developed in our research group. With this tool, 1P could form nanoparticles 1-NPs under the catalysis of ALP and thus could be used to enhance PA imaging both in vitro and in vivo.

Photoacoustic probe of targeting intracellular Staphylococcus aureus infection with signal-enhanced by self-assembly.

In this chapter, we introduce the photoacoustic imaging probe in detecting the infection of bacterial in host cells. We give detailed protocols for the synthesis and confirmation of a PA probe which response to caspase-1 and then self-assembly to enhance the signal. With this PA contrast agent, we may provide a new approach for an intracellular bacterial infection with the selective and sensitive diagnosis.

Synthesis of deep red emitting Cu–In–Zn–Se/ZnSe quantum dots for dual-modal fluorescence and photoacoustic imaging

CuInSe2 quantum dots (QDs) are one of the most important Cd-free fluorescent probes; they usually exhibited low fluorescence intensity, suggesting that a considerable amount of absorbed photon energy was lost as heat. In this study we aimed to improve the fluorescence intensity of CuInSe2 QDs and investigate their photoacoustic (PA) signal resulting from the heat dissipation, which was previously rarely reported. Cu–In–Zn–Se/ZnSe QDs were synthesized by adopting two strategies of Zn doping and ZnSe shell growth. It was found that there was an upper limit for Zn concentration beyond which the fluorescence intensity began to decrease. In addition, a blue shift of the emission peak of Cu–In–Zn–Se/ZnSe QDs was observed at high concentrations of ZnSe precursor due to the diffusion of excessive Zn. To prepare the dual-modal fluorescence and PA imaging probe, poly(maleic anhydride-alt-1-octadecene) (PMAO) modified with polyethylene glycol (PEG) was coated on the QDs, which led to a slight reduction in fluorescence. Cellular labeling on HeLa cells was performed to demonstrate the utility of these probes for fluorescence imaging. We further studied the in vitro PA imaging capabilities of the Cu–In–Zn–Se/ZnSe/PMAO-g-PEG nanoparticles, which showed a distinct PA signal beyond 1.0 mg ml−1. The current work demonstrated that a moderate amount of Zn doping is necessary for enhancing fluorescence and there is a limit beyond which the fluorescence will be diminished. We also demonstrated the proof of concept that Cu–In–Zn–Se/ZnSe QDs are able to serve as a potential PA imaging contrast agent.

Croconaine-Based Polymer Particles as Contrast Agents for Photoacoustic Imaging.

In the development and optimization of imaging methods, photoacoustic imaging (PAI) has become a powerful tool for preclinical biomedical diagnosis and detection of cancer. PAI probes can improve contrast and help identify pathogenic tissue. Such contrast agents must meet several requirements: they need to be biocompatible, and absorb strongly in the near-infrared (NIR) range, while relaxing the photoexcited state thermally and not radiatively. In this work, polymer nanoparticles are produced with croconaine as a monomer unit. Small molecular croconaine dyes are known to act as efficient pigments, which do not show photoluminescence. Here, for the first time croconaine copolymer nanoparticles are produced from croconic acid and a range of aromatic diamines. Following a dispersion polymerization protocol, this approach yields monodisperse particles of adjustable size. All synthesized polymers exhibit broad absorption within the NIR spectrum and therefore represent suitable candidates as contrast agents for PAI. The optical properties of these polymer particles are discussed with respect to the relation between particle size and outstanding photoacoustic performance. Biocompatibility of the polymer particles is demonstrated in cell viability experiments.

Enhanced optical imaging properties of lipid nanocapsules as vehicles for fluorescent conjugated polymers

Conjugated polymer nanoparticles (CPNs) have emerged as highly photostable probes for optical and photoacoustic imaging. However, the aggregation of conjugated polymer (CP) molecules upon nanoparticle formation is associated with fluorescence quenching, poor yields and mutable particle sizes. This study investigated whether the CP encapsulation within the liquid midchain triglyceride (MCT) core of lipid nanocapsules (LNCs) may achieve reduced packing of CP chains leading to a stable system with enhanced optical features. The red- and near infrared-emitting CPs, CN-PPV and PCPDTBT, showed precipitation and aggregation-induced quenching with concentrations >~25 µg/mL in MCT alone. Despite this, CP encapsulation within LNCs abolished quenching at concentrations up to 1500 µg/mL. PCPDTBT-LNCs exhibited a quantum yield of 2.8% and a higher signal:background ratio in an optical imaging phantom compared to literature reports of PCPDTBT encapsulated in PEG-PLGA nanoparticles. In contrast, PCPDTBT-LNCs had slightly lower photoacoustic amplitudes than reported PEG-PLGA systems. CP-LNCs were also stable in size (32 ± 0.7 nm) and photoluminescence over 21 days at 4 °C, 25 °C and 37 °C. In summary, encapsulation of CP within the liquid core of lipid nanocapsules enhances the optical properties of fluorescent CP.

In Vivo Dual-Modal Photoacoustic and Ultrasound Imaging of Sentinel Lymph Nodes Using a Solid-State Dye Laser System.

Photoacoustic imaging (PAI) is being actively investigated as a non-invasive and non-radioactive imaging technique for sentinel lymph node (SLN) biopsy. By taking advantage of optical and ultrasound imaging, PAI probes SLNs non-invasively with methylene blue (MB) in both live animals and breast cancer patients. However, these PAI systems have limitations for widespread use in clinics and commercial marketplaces because the lasers used by the PAI systems, e.g., tunable liquid dye laser systems and optical parametric oscillator (OPO) lasers, are bulky in size, not economical, and use risky flammable and toxic liquid dyes. To overcome these limitations, we are proposing a novel dual-modal photoacoustic and ultrasound imaging system based on a solid-state dye laser (SD-PAUSI), which is compact, convenient, and carries far less risk of flammability and toxicity. Using a solid-state dye handpiece that generates 650-nm wavelength, we successfully imaged the MB tube positioned deeply (~3.9 cm) in chicken breast tissue. The SLNs were also photoacoustically detected in the in vivo rats beneath a 2.2-cm-thick layer of chicken breast, which is deeper than the typical depth of SLNs in humans (1.2 ± 0.5 cm). Furthermore, we showed the multispectral capability of the PAI by switching the dye handpiece, in which the MB-dyed SLN was selectively highlighted from the surrounding vasculature. These results demonstrated the great potential of the SD-PAUSI as an easy but effective modality for SLN detection.

Controllable Cleavage of C-N Bond-Based Fluorescent and Photoacoustic Dual-Modal Probes for the Detection of H2S in Living Mice.

Hydrogen sulfide (H2S) has been recognized to influence a wide range of physiological and pathological processes. Its underlying molecular events, however, are still poorly understood. An activatable H2S probe for photoacoustic (PA) imaging is desirable to further explore the role of H2S in vivo. Nevertheless, only a few activatable PA probes for H2S detection have been reported. In particular, examples of dual-modal H2S probes with the combined advantages of fluorescence (high sensitivity and resolution) and PA imaging (deep penetration) are very rare. Herein the controllable cleavage of the C-N bond in nitrobenzoxadiazole (NBD) amine derivatives by H2S is presented for the first time. The cleavage reactivity was found to be accelerated by the introduction of an electron-withdrawing group. Through this strategy, a series of fluorescent and PA dual-modal probes (1-3) were developed for H2S detection. Among them, probe 3 shows a high fluorescence on-off response rate (k2 = 4.04 M-1 s-1) and excellent selectivity for H2S over other biothiols. Moreover, probe 3 can also work as an activatable PA H2S probe because of the significant shift of its absorption peak from 468 to 532 nm in the H2S reaction. Importantly, probe 3 demonstrates its capability for fluorescence and PA imaging of H2S in living cells and mice. These results indicate that the controllable cleavage of the C-N bond can serve as an efficient strategy for designing fluorescent and PA dual-modal H2S probes.

Application of functionalized carbon dots in detection, diagnostic, disease treatment, and desalination: a review

Particularly in recent years, fluorescence carbon dots (CDs) were in the central point of scientist’s focus. However, some difficulties may exist concerning CDs. For example, their targeting by other analytes usually goes wrong due to the absence of specific recognition groups on the surface of CDs. Furthermore, due to low quantum yield, they cannot be conducive to bioimaging. Additionally, they do not have biological functionality that originated from poor interaction with biological agents. For improvement of the luminescence properties and extending of new applications for CDs, the functionalisation of CDs has attracted many interests. Functionalised CDs have promising applications. The present paper has reviewed different published reports considering applications of functionalised CDs. In this comprehensive article, first the functionalisation mechanisms of CDs are discussed. Then, the functionalised CDs for detection applications including biochemical and chemical detections are reviewed. After that, functionalised CDs for diagnostic applications are reviewed. In this part functionalised CDs for fluorescence imaging probes, functionalised CDs for surface-enhanced Raman scattering (SERS) imaging and functionalised CDs for photoacoustic imaging probes are discussed. Later, functionalised CDs for disease treatment including drug delivery and photothermal therapy applications are reviewed to elucidate the importance of their function in cancer treatment. Finally, the applications of functionalised CDs for water desalination are reviewed.

Photoacoustic Imaging Probes Based on Tetrapyrroles and Related Compounds.

Photoacoustic imaging (PAI) is a rapidly evolving field in molecular imaging that enables imaging in the depths of ultrasound and with the sensitivity of optical modalities. PAI bases on the photoexcitation of a chromophore, which converts the absorbed light into thermal energy, causing an acoustic pressure wave that can be captured with ultrasound transducers, in generating an image. For in vivo imaging, chromophores strongly absorbing in the near-infrared range (NIR; > 680 nm) are required. As tetrapyrroles have a long history in biomedical applications, novel tetrapyrroles and inspired mimics have been pursued as potentially suitable contrast agents for PAI. The goal of this review is to summarize the current state of the art in PAI applications using tetrapyrroles and related macrocycles inspired by it, highlighting those compounds exhibiting strong NIR-absorption. Furthermore, we discuss the current developments of other absorbers for in vivo photoacoustic (PA) applications.

A dual-modal probe for NIR fluorogenic and ratiometric photoacoustic imaging of Cys/Hcy in vivo

Biothiols, such as cysteine (Cys) and homocysteine (Hcy), play vital roles in biological homeostasis and are closely related to various pathological and physiological processes in the living systems. Therefore, the in vivo detection of biothiols is of great importance for early diagnosis of diseases and assessment of disease progression. In this work, we developed a near-infrared (NIR) fluorescence and photoacoustic dual-modal molecular probe ( NIR-S ) that can be specifically activated by Cys or Hcy. The aryl-thioether substituted cyanine probe can undergo nucleophilic substitution and Smiles rearrangement reaction, resulting in specific turn-on NIR fluorescence and ratiometric photoacoustic responses for Hcy/Cys. Thus, NIR-S not only realizes the specific NIR fluorescence and photoacoustic dual mode imaging to detect Hcy/Cys in solution, but also can be applied to living cells and mice to detect Hcy/Cys. This work provided a practical tool to detect Hcy/Cys levels in vivo , which would be beneficial for the early diagnosis and progress of diseases.

Three-dimensional view of out-of-plane artifacts in photoacoustic imaging using a laser-integrated linear-transducer-array probe

Research on photoacoustic imaging (PAI) using a handheld integrated photoacoustic probe has been a recent focus of clinical translation of this imaging technique. One of the remaining challenges is the occurrence of out-of-plane artifacts (OPAs) in such a probe. Previously, we proposed a method to identify and remove OPAs by axially displacing the transducer array. Here we show that besides the benefit of removing OPAs from the imaging plane, the proposed method can provide a three-dimensional (3D) view of the OPAs. In this work, we present a 3D reconstruction method using axial transducer array displacement. By axially displacing the transducer array, out-of-plane absorbers can be three-dimensionally visualized at an elevation distance of up to the acquired imaging depth. Additionally, OPAs in the in-plane image are significantly reduced. We experimentally demonstrate the method with phantom and in vivo experiments using an integrated PAI probe. We also compare the method with elevational transducer array displacement and take into account the sensitivity of the transducer array in the 3D reconstruction.

Maltotriose-based probes for fluorescence and photoacoustic imaging of bacterial infections

Currently, there are no non-invasive tools to accurately diagnose wound and surgical site infections before they become systemic or cause significant anatomical damage. Fluorescence and photoacoustic imaging are cost-effective imaging modalities that can be used to noninvasively diagnose bacterial infections when paired with a molecularly targeted infection imaging agent. Here, we develop a fluorescent derivative of maltotriose (Cy7-1-maltotriose), which is shown to be taken up in a variety of gram-positive and gram-negative bacterial strains in vitro. In vivo fluorescence and photoacoustic imaging studies highlight the ability of this probe to detect infection, assess infection burden, and visualize the effectiveness of antibiotic treatment in E. coli-induced myositis and a clinically relevant S. aureus wound infection murine model. In addition, we show that maltotriose is an ideal scaffold for infection imaging agents encompassing better pharmacokinetic properties and in vivo stability than other maltodextrins (e.g. maltohexose).

Conjugated polymers as nanoparticle probes for fluorescence and photoacoustic imaging.

Bioimaging enables the visualisation of biological processes at the microscopic and macroscopic levels, finding applications from cellular tracking to whole body scanning for diagnosis purposes. The different techniques developed to acquire images make use of most radiation types of the electromagnetic spectrum. Recently, there has been interest in non-ionising radiation imaging techniques that can offer improved detection or additional information about biological processes, and fluorescence and photoacoustic imaging have become an eminent field. Conjugated polymers are versatile materials for bioimaging due to their tailored absorption and emission spectra and applications in both fluorescence and photoacoustic imaging. This review gives an overview on bioimaging techniques, with a special focus on conjugated polymer nanoparticles (CPNs), the different types of nanoparticle chemistries published and their preclinical safety assessment.

In situ visualization of peroxisomal viscosity in the liver of mice with non-alcoholic fatty liver disease by near-infrared fluorescence and photoacoustic imaging.

Non-alcoholic fatty liver disease (NAFLD) can gradually develop into hepatic failure, and early diagnosis is crucial to improve treatment efficiency. The occurrence of NAFLD is closely related to lipid metabolism. Peroxisomes act as the first and main site for lipid metabolism in the hepatocytes, so abnormal lipid metabolism might directly affect peroxisomal viscosity. Herein, we developed a new near-infrared fluorescence (NIRF) and photoacoustic (PA) imaging probe (PV-1) for the real-time visualization of peroxisomal viscosity in vivo. This PV-1 encompasses the malononitrile group as the rotor, which emits strong NIRF (at 705 nm) and PA (at 680 nm) signals when rotation is hindered as viscosity increases. Through dual-mode imaging, we discovered distinctly higher viscosity in the liver of NAFLD mice for the first time. We further found the remarkable amelioration of NAFLD upon treatment with N-acetylcysteine (NAC). Therefore, we anticipate that the PV-1 imaging method is promising for the early diagnosis and prognostic evaluation of NAFLD.

An Activatable Cancer-Targeted Hydrogen Peroxide Probe for Photoacoustic and Fluorescence Imaging.

Reactive oxygen species play an important role in cancer, however, their promiscuous reactivity, low abundance, and short-lived nature limit our ability to study them in real time in living subjects with conventional noninvasive imaging methods. Photoacoustic imaging is an emerging modality for in vivo visualization of molecular processes with deep tissue penetration and high spatiotemporal resolution. Here, we describe the design and synthesis of a targeted, activatable probe for photoacoustic imaging, which is responsive to one of the major and abundant reactive oxygen species, hydrogen peroxide (H2O2). This bifunctional probe, which is also detectable with fluorescence imaging, is composed of a heptamethine carbocyanine dye scaffold for signal generation, a 2-deoxyglucose cancer localization moiety, and a boronic ester functionality that specifically detects and reacts to H2O2. The optical properties of the probe were characterized using absorption, fluorescence, and photoacoustic measurements; upon addition of pathophysiologic H2O2 concentrations, a clear increase in fluorescence and red-shift of the absorption and photoacoustic spectra were observed. Studies performed in vitro showed no significant toxicity and specific uptake of the probe into the cytosol in breast cancer cell lines. Importantly, intravenous injection of the probe led to targeted uptake and accumulation in solid tumors, which enabled noninvasive photoacoustic and fluorescence imaging of H2O2. In conclusion, the reported probe shows promise for the in vivo visualization of hydrogen peroxide. SIGNIFICANCE: This study presents the first activatable and cancer-targeted hydrogen peroxide probe for photoacoustic molecular imaging, paving the way for visualization of hydrogen peroxide at high spatiotemporal resolution in living subjects.Graphical Abstract: http://cancerres.aacrjournals.org/content/canres/79/20/5407/F1.large.jpg.

Ultrasound-Responsive Conversion of Microbubbles to Nanoparticles to Enable Background-Free in Vivo Photoacoustic Imaging.

Photoacoustic (PA) imaging based on the photon-to-ultrasound conversion allows the imaging of optical absorbers in deep tissues with high spatial resolution. However, the inherent optical absorbance of biomolecules (e.g., hemoglobin, melanin, etc.) would show up as tissue background signals to interfere with signals from the contrast agent during in vivo PA imaging, limiting the imaging sensitivity. Herein, an ultrasound (US)-responsive PA imaging probe based on microbubbles (MBs) containing gold nanoparticles (Au NPs) is designed for in vivo "background-free" PA imaging. The obtained Au@lip MBs with separated Au NPs decorated within the lipid shell of MBs show low PA signals under near-infrared (NIR) excitation. Interestingly, under exposure to US pulses, those Au@lip MBs would burst to form nanoscale aggregates of Au@lip NPs, which exhibit significantly enhanced NIR PA signals due to their red-shifted surface plasmon resonance. Therefore, by subtracting the PA image captured pre-US burst from that captured post-US burst, the tissue background PA signals could be deducted to enable background-free PA imaging with high sensitivities as demonstrated by multiple ex vivo and in vivo experiments. This work presents a simple yet effective strategy to deduct background signals during PA imaging, which is promising for accurate PA detection of targets in tissues with a strong background.

Polymer-based activatable optical probes for tumor fluorescence and photoacoustic imaging.

Optical imaging including fluorescence imaging and photoacoustic imaging have been widely employed in early and accurate diagnosis of cancer. Compared to the "always on" optical probes, the molecular probes that could emit their signal in response to the tumor microenvironment exhibit the low background noise and high signal-to-background ratio, allowing sensitive and accurate cancer diagnosis. Polymer-based activatable optical probes display the advantages of improved water solubility, good photostability, extended blood circulation time, and easy functionalization, which enable them to accumulate in tumor for early and accurate diagnosis. This review focuses on recent advances in the development of polymer-based activatable optical probes for tumor fluorescence and PA imaging. The designs of polymer-based optical probes are first discussed. Then the applications for tumor fluorescence and PA imaging using pH, hypoxia, reactive oxygen and nitrogen species, and enzymes responsive polymer-based optical probes are discussed in details. At last, the present challenges and perspectives of polymer-based activatable optical probes to further advance them into the clinical application are also suggested. This article is categorized under: Diagnostic Tools > In vivo Nanodiagnostics and Imaging Diagnostic Tools > Biosensing.