RESUMO
To identify the vascular alteration by photodynamic therapy (PDT), the utilization of high-resolution, high-speed, and wide-field photoacoustic microscopy (PAM) has gained enormous interest. The rapid changes in vasculature during PDT treatment and monitoring of tumor tissue activation in the orthotopic pancreatic cancer model have received limited attention in previous studies. Here, a fully two-axes waterproof galvanometer scanner-based photoacoustic microscopy (WGS-PAM) system was developed for in vivo monitoring of dynamic variations in micro blood vessels due to PDT in an orthotopic pancreatic cancer mouse model. The photosensitizer (PS), Chlorin e6 (Ce6), was utilized to activate antitumor reactions in response to the irradiation of a 660 nm light source. Microvasculatures of angiogenesis tissue were visualized on a 40 mm2 area using the WGS-PAM system at 30 min intervals for 3 h after the PDT treatment. The decline in vascular intensity was observed at 24.5% along with a 32.4% reduction of the vascular density at 3 h post-PDT by the analysis of PAM images. The anti-vascularization effect was also identified with fluorescent imaging. Moreover, Ce6-PDT increased apoptotic and necrotic markers while decreasing vascular endothelial growth factor (VEGF) expression in MIA PaCa-2 and BxPC-3 pancreatic cancer cell lines. The approach of the WGS-PAM system shows the potential to investigate PDT effects on the mechanism of angiographic dynamics with high-resolution wide-field imaging modalities.
Assuntos
Clorofilídeos , Neoplasias Pancreáticas , Fotoquimioterapia , Porfirinas , Camundongos , Animais , Fotoquimioterapia/métodos , Microscopia , Fator A de Crescimento do Endotélio Vascular/uso terapêutico , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/uso terapêutico , Neoplasias Pancreáticas/diagnóstico por imagem , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Pancreáticas/patologia , Linhagem Celular Tumoral , Porfirinas/farmacologia , Porfirinas/uso terapêuticoRESUMO
In this paper, a portable photoacoustic microscopy (PAM) system is proposed based on a large stroke electrothermal micromirror to achieve high resolution and fast imaging. The crucial micromirror in the system realizes a precise and efficient 2-axis control. Two different designs of electrothermal actuators with "O" and "Z" shape are evenly located around the four directions of mirror plate. With a symmetrical structure, the actuator realized single direction drive only. The finite element modelling of both two proposed micromirror has realized a large displacement over 550 µm and the scan angle over ±30.43° at 0-10 V DC excitation. In addition, the steady-state and transient-state response show a high linearity and quick response respectively, which can contribute to a fast and stable imaging. Using the Linescan model, the system achieves an effective imaging area of 1 mm × 3 mm in 14 s and 1 mm × 4 mm in 12 s for the "O" and "Z" types, respectively. The proposed PAM systems have advantages in image resolution and control accuracy, indicating a significant potential in the field of facial angiography.
Assuntos
Técnicas Fotoacústicas , Técnicas Fotoacústicas/métodos , Microscopia/métodos , Angiografia , Análise EspectralRESUMO
Photoacoustic imaging has emerged as a promising biomedical imaging technique that enables visualization of the optical absorption characteristics of biological tissues in vivo. Among the different photoacoustic imaging system configurations, optical-resolution photoacoustic microscopy stands out by providing high spatial resolution using a tightly focused laser beam, which is typically transmitted through optical fibers. Achieving high-quality images depends significantly on optical fluence, which is directly proportional to the signal-to-noise ratio. Hence, optimizing the laser-fiber coupling is critical. Conventional coupling systems require manual adjustment of the optical path to direct the laser beam into the fiber, which is a repetitive and time-consuming process. In this study, we propose an automated laser-fiber coupling module that optimizes laser delivery and minimizes the need for manual intervention. By incorporating a motor-mounted mirror holder and proportional derivative control, we successfully achieved efficient and robust laser delivery. The performance of the proposed system was evaluated using a leaf-skeleton phantom in vitro and a human finger in vivo, resulting in high-quality photoacoustic images. This innovation has the potential to significantly enhance the quality and efficiency of optical-resolution photoacoustic microscopy.
RESUMO
Choroidal neovascularization (CNV) in young rabbits has been shown to have a rapid, robust response after treatment with bevacizumab, an anti-vascular endothelial growth factor (VEGF) medication. This investigation evaluates an age differential response to bevacizumab in older populations of rabbits using multimodal high resolution molecular imaging. Young (4 months old) and life span (14 months old) rabbits were given subretinal injections of Matrigel and VEGF to produce CNV. All CNV rabbit models were then treated with a bevacizumab intravitreal injection. Rabbits were then monitored longitudinally using photoacoustic microscopy (PAM), optical coherence tomography (OCT), color photography, and fluorescence imaging. Chain-like gold nanoparticle clusters (CGNP) conjugated with tripeptide arginylglycylaspartic acid (RGD) was injected intravenously for molecular imaging. Robust CNV developed in both young and old rabbits. After intravitreal bevacizumab injection, fluorescence signals were markedly decreased 90.13% in the young group. In contrast, old rabbit CNV area decreased by only 10.56% post-bevacizumab treatment. OCT images confirmed a rapid decrease of CNV in the young group. CGNPs demonstrated high PAM signal in old rabbits and minimal PAM signal in young rabbits after bevacizumab, indicating CNV regression. There is a significant difference in response to intravitreal bevacizumab treatment between young and old rabbits with CNV which can be monitored with multimodal molecular imaging. Old rabbits demonstrate significant persistent disease activity. This represents the first large eye model of persistent disease activity of CNV and could serve as the foundation for future investigations into the mechanism of persistent disease activity and the development of novel therapies.
Assuntos
Neovascularização de Coroide , Nanopartículas Metálicas , Inibidores da Angiogênese/uso terapêutico , Animais , Anticorpos Monoclonais Humanizados/uso terapêutico , Bevacizumab/uso terapêutico , Neovascularização de Coroide/tratamento farmacológico , Fatores de Crescimento Endotelial/uso terapêutico , Angiofluoresceinografia , Ouro , Injeções Intravítreas , Oligopeptídeos/uso terapêutico , Coelhos , Tomografia de Coerência Óptica/métodos , Fator A de Crescimento do Endotélio Vascular , Acuidade VisualRESUMO
Despite all the expectations for photoacoustic endoscopy (PAE), there are still several technical issues that must be resolved before the technique can be successfully translated into clinics. Among these, electromagnetic interference (EMI) noise, in addition to the limited signal-to-noise ratio (SNR), have hindered the rapid development of related technologies. Unlike endoscopic ultrasound, in which the SNR can be increased by simply applying a higher pulsing voltage, there is a fundamental limitation in leveraging the SNR of PAE signals because they are mostly determined by the optical pulse energy applied, which must be within the safety limits. Moreover, a typical PAE hardware situation requires a wide separation between the ultrasonic sensor and the amplifier, meaning that it is not easy to build an ideal PAE system that would be unaffected by EMI noise. With the intention of expediting the progress of related research, in this study, we investigated the feasibility of deep-learning-based EMI noise removal involved in PAE image processing. In particular, we selected four fully convolutional neural network architectures, U-Net, Segnet, FCN-16s, and FCN-8s, and observed that a modified U-Net architecture outperformed the other architectures in the EMI noise removal. Classical filter methods were also compared to confirm the superiority of the deep-learning-based approach. Still, it was by the U-Net architecture that we were able to successfully produce a denoised 3D vasculature map that could even depict the mesh-like capillary networks distributed in the wall of a rat colorectum. As the development of a low-cost laser diode or LED-based photoacoustic tomography (PAT) system is now emerging as one of the important topics in PAT, we expect that the presented AI strategy for the removal of EMI noise could be broadly applicable to many areas of PAT, in which the ability to apply a hardware-based prevention method is limited and thus EMI noise appears more prominently due to poor SNR.
Assuntos
Aprendizado Profundo , Algoritmos , Animais , Fenômenos Eletromagnéticos , Endoscopia , Processamento de Imagem Assistida por Computador/métodos , RatosRESUMO
Revealing the structural and functional change of microvasculature is essential to match vascular response with neuronal activities in the investigation of neurovascular coupling. The increasing use of rhesus models in fundamental and clinical studies of neurovascular coupling presents an emerging need for a new imaging modality. Here we report a structural and functional cerebral vascular study of rhesus monkeys using an ultrafast, portable, and high resolution photoacoustic microscopic system with a long working distance and a special scanning mechanism to eliminate the relative displacement between the imaging interface and samples. We derived the structural and functional response of the cerebral vasculature to the alternating normoxic and hypoxic conditions by calculating the vascular diameter and functional connectivity. Both vasodilatation and vasoconstriction were observed in hypoxia. In addition to the change of vascular diameter, the decrease of functional connectivity is also an important phenomenon induced by the reduction of oxygen ventilatory. These results suggest that photoacoustic microscopy is a promising method to study the neurovascular coupling and cerebral vascular diseases due to the advanced features of high spatiotemporal resolution, excellent sensitivity to hemoglobin, and label-free imaging capability of observing hemodynamics.
Assuntos
Córtex Cerebral/diagnóstico por imagem , Circulação Cerebrovascular/fisiologia , Hemodinâmica/fisiologia , Microscopia/métodos , Animais , Macaca mulatta , Acoplamento Neurovascular/fisiologia , Técnicas Fotoacústicas , Vasoconstrição/fisiologia , Vasodilatação/fisiologiaRESUMO
Microcirculatory changes and oxidative stress have long been associated with acute kidney injury. Despite substantial progress made by two-photon microscopy of microvascular responses to acute kidney injury in rodent models, little is known about the underlying changes in blood oxygen delivery and tissue oxygen metabolism. To fill this gap, we developed a label-free kidney imaging technique based on photoacoustic microscopy, which enables simultaneous quantification of hemoglobin concentration, oxygen saturation of hemoglobin, and blood flow in peritubular capillaries in vivo. Based on these microvascular parameters, microregional oxygen metabolism was quantified. We demonstrated the utility of this technique by studying kidney hemodynamic and oxygen-metabolic responses to acute kidney injury in mice subject to lipopolysaccharide-induced sepsis. Dynamic photoacoustic microscopy of the peritubular capillary function and tissue oxygen metabolism revealed that sepsis induced an acute and significant reduction in peritubular capillary oxygen saturation of hemoglobin, concomitant with a marked reduction in kidney ATP levels and contrasted with nominal changes in peritubular capillary flow and plasma creatinine. Thus, our technique opens new opportunities to study microvascular and metabolic dysfunction in acute and chronic kidney diseases.
Assuntos
Capilares , Microscopia , Animais , Rim , Camundongos , Microcirculação , OxigênioRESUMO
Photoacoustic microscopy (PAM) has significant potential as a promising diagnostic method for eye diseases and can provide anatomic and functional information of the retinal and choroidal vasculature. However, there are no FDA-approved PAM systems for ophthalmic imaging. In this study, a comprehensive safety evaluation was performed to evaluate the safety of PAM retinal imaging and whether PAM causes damage to retinal structure or function in rabbit eyes. 12 Dutch-Belted pigmented rabbits received photoacoustic imaging to 57% of the retinal surface area with a laser energy of 5% of the ANSI safety limit for five consecutive days and followed before imaging and 3 days, 1, 2, 3, and 4 weeks post imaging. Retinal morphologic analyses using slit lamp examination, fundus photography, red free, FA, FAF, ICGA, and OCT showed no retinal hemorrhage, edema, detachment, vascular abnormalities, or pigmentary abnormalities in the retina or choroid after PAM imaging. Full-field ERG analysis showed no significant difference in scotopic or photopic a- and b-wave amplitudes or implicit times between the control and experimental eyes over time (n = 6, P values > 0.05). Retinal ultrastructural evaluation using TEM showed normal structure of organelles and nuclei, and no significant loss of cells after PAM. TUNEL assay showed no evidence of cells apoptosis in retina. Retinal histopathology indicated that the architecture and thickness of the retinal layers was well preserved in all experimental eyes. A positive control at 500% of the ANSI limit demonstrated significant damage. The comprehensive retinal safety evaluation demonstrated no damage to retinal structure or function for 4 weeks after PAM imaging in rabbits.
Assuntos
Microscopia Acústica/métodos , Técnicas Fotoacústicas/métodos , Retina/diagnóstico por imagem , Animais , Modelos Animais , Coelhos , Reprodutibilidade dos Testes , Vasos Retinianos/diagnóstico por imagemRESUMO
Carfilzomib is mainly used to treat multiple myeloma. Several side effects have been reported in patients treated with carfilzomib, especially those associated with cardiovascular events, such as hypertension, congestive heart failure, and coronary artery disease. However, the side effects, especially the manifestation of cardiovascular events through capillaries, have not been fully investigated. Here, we performed a pilot experiment to monitor peripheral vascular dynamics in a mouse ear under the effects of carfilzomib using a quantitative photoacoustic vascular evaluation method. Before and after injecting the carfilzomib, bortezomib, and PBS solutions, we acquired high-resolution three-dimensional PAM data of the peripheral vasculature of the mouse ear during each experiment for 10 h. Then, the PAM maximum amplitude projection (MAP) images and five quantitative vascular parameters, i.e., photoacoustic (PA) signal, diameter, density, length fraction, and fractal dimension, were estimated. Quantitative results showed that carfilzomib induces a strong effect on the peripheral vascular system through a significant increase in all vascular parameters up to 50%, especially during the first 30 min after injection. Meanwhile, bortezomib and PBS do not have much impact on the peripheral vascular system. This pilot study verified PAM as a comprehensive method to investigate peripheral vasculature, along with the effects of carfilzomib. Therefore, we expect that PAM may be useful to predict cardiovascular events caused by carfilzomib.
Assuntos
Mieloma Múltiplo , Oligopeptídeos , Animais , Bortezomib/toxicidade , Humanos , Camundongos , Oligopeptídeos/farmacologia , Oligopeptídeos/toxicidade , Projetos PilotoRESUMO
Vascular damage is one of the therapeutic mechanisms of photodynamic therapy (PDT). In particular, short-term PDT treatments can effectively destroy malignant lesions while minimizing damage to nonmalignant tissue. In this study, we investigate the feasibility of label-free quantitative photoacoustic microscopy (PAM) for monitoring the vasculature changes under the effect of PDT in mouse ear melanoma tumors. In particular, quantitative vasculature evaluation was conducted based on Hessian filter segmentation. Three-dimensional morphological PAM and depth-resolved images before and after PDT treatment were acquired. In addition, five quantitative vasculature parameters, including the PA signal, vessel diameter, vessel density, perfused vessel density, and vessel complexity, were analyzed to evaluate the influence of PDT on four different areas: Two melanoma tumors, and control and normal vessel areas. The quantitative and qualitative results successfully demonstrated the potential of the proposed PAM-based quantitative approach to evaluate the effectiveness of the PDT method.
Assuntos
Melanoma , Técnicas Fotoacústicas , Fotoquimioterapia , Animais , Melanoma/diagnóstico por imagem , Melanoma/tratamento farmacológico , Camundongos , Microscopia , Fármacos Fotossensibilizantes , Análise EspectralRESUMO
Photoacoustic imaging (PAI) is a rapidly developing hybrid biomedical imaging technology, which is capable of providing structural and functional information of biological tissues. Due to inevitable motion of the imaging object, such as respiration, heartbeat or eye rotation, motion artifacts are observed in the reconstructed images, which reduce the imaging resolution and increase the difficulty of obtaining high-quality images. This paper summarizes current methods for correcting and compensating motion artifacts in photoacoustic microscopy (PAM) and photoacoustic tomography (PAT), discusses their advantages and limits and forecasts possible future work.
Assuntos
Artefatos , Técnicas Fotoacústicas , Microscopia , Movimento (Física) , Tomografia Computadorizada por Raios XRESUMO
PURPOSE: To demonstrate the value of the laser-scanning optical-resolution (LSOR)-photoacoustic (PA) microscopy (PAM) system and the conventional multimodal imaging techniques in the evaluation of laser-induced retinal injury and choroidal neovascularization (CNV) in rats. METHODS: Different degrees of retinal injury were induced using laser photocoagulation. We compared the LSOR-PAM system with conventional imaging techniques in evaluating retinal injury with or without CNV. Six additional rats, treated with an anti-VEGF antibody or immunoglobulin G immediately after photocoagulation, were imaged 7 and 14 days after injection, and CNV lesion areas were compared. RESULTS: In the retinal injury model, fundus autofluorescence showed well-defined hyperreflection, while the lesion displayed abundant PA signals demonstrating nonuniform melanin distribution in retinal pigment epithelium (RPE). RPE was detected with higher contrast in the PAM B-scan image than optical coherence tomography (OCT). Additionally, the CNV lesion was present with multiple PA signal intensities which distinctly characterized the location and area of CNV as found in fundus fluorescein angiography. Furthermore, the decreased PA signals extending from the CNV lesion were similar to those of the vascular bud in ex vivo imaging, which was invisible in other in vivo images. When treated with anti-VEGF agents, statistically significant differences can be demonstrated by PAM similar to other modalities. CONCLUSIONS: LSOR-PAM can detect the melanin distribution of RPE in laser-induced retinal injury and CNV in rats. PAM imaging provides a potential new tool to evaluate the vitality and functionality of RPE in vivo as well as to monitor the development and treatment of CNV.
Assuntos
Neovascularização de Coroide/diagnóstico , Microscopia Acústica/métodos , Epitélio Pigmentado da Retina/patologia , Animais , Neovascularização de Coroide/etiologia , Modelos Animais de Doenças , Fotocoagulação a Laser/efeitos adversos , Masculino , Ratos , Ratos Endogâmicos BNRESUMO
In this study, a photoacoustic microscopy (PAM) system based on a multifocal point (MFP) transducer was fabricated to produce a large depth-of-field tissue image. The customized MFP transducer has seven focal points, distributed along with the transducer's axis, fabricated by separate spherically-focused surfaces. These surfaces generate distinct focal zones that are overlapped to extend the depth-of-field. This design allows extending the focal zone of 10 mm for the 11 MHz MFP transducer, which is a great improvement over the 0.48 mm focal zone of the 11 MHz single focal point (SFP) transducer. The PAM image penetration depths of a chicken-hemoglobin phantom using SFP and MFP transducers were measured as 5 mm and 8 mm, respectively. The significant increase in the PAM image-based penetration depth of the chicken-hemoglobin phantom was a result of using the customized MFP transducer.
Assuntos
Microscopia/métodos , Técnicas Fotoacústicas , Transdutores , Animais , Galinhas , Desenho de Equipamento , Hemoglobinas/análise , Processamento de Imagem Assistida por Computador , Carne/análise , Microscopia/instrumentação , UltrassonografiaRESUMO
Photoacoustic microscopy is a novel, non-ionizing, non-invasive imaging technology that evaluates tissue absorption of short-pulsed light through the sound waves emitted by the tissue and has numerous biomedical applications. In this study, a custom-built multimodal imaging system, including photoacoustic microscopy (PAM) and optical coherence tomography (OCT), has been developed to evaluate choroidal vascular occlusion (CVO). CVO was performed on three living rabbits using laser photocoagulation. Longitudinal imaging of CVO was obtained using multiple imaging tools such as color fundus photography, fluorescein angiography, indocyanine green angiography (ICGA), OCT, and PAM. PAM images were acquired at different wavelengths, ranging from 532 to 700 nm. The results demonstrate that the CVO was clearly observed on PAM in both two dimensions (2D) and 3D with high resolution longitudinally over 28 days. In addition, the location and margin of the CVO were distinguished from the surrounding choroidal vasculature after the injection of ICG contrast agent. PAM imaging was achieved using a laser energy of approximately 80 nJ, which is about half of the American National Standards Institute safety limit. The proposed imaging technique may provide a potential tool for the evaluation of different chorioretinal vascular disease pathogeneses and other biological studies.
Assuntos
Microscopia Acústica/métodos , Imagem Multimodal/métodos , Tomografia de Coerência Óptica/métodos , Animais , Corioide/irrigação sanguínea , Corioide/diagnóstico por imagem , Angiofluoresceinografia/métodos , Lasers , Microscopia/métodos , Imagem Multimodal/instrumentação , Técnicas Fotoacústicas/métodos , Coelhos , Retina/diagnóstico por imagem , Retina/metabolismo , Análise EspectralRESUMO
Cerebral small vessel disease has been linked to cognitive, psychiatric and physical disabilities, especially in the elderly. However, the underlying pathophysiology remains incompletely understood, largely due to the limited accessibility of these small vessels in the live brain. Here, we report an intravital imaging and analysis platform for high-resolution, quantitative and comprehensive characterization of pathological alterations in the mouse cerebral microvasculature. By exploiting multi-parametric photoacoustic microscopy (PAM), microvascular structure, blood perfusion, oxygenation and flow were imaged in the awake brain. With the aid of vessel segmentation, these structural and functional parameters were extracted at the single-microvessel level, from which vascular density, tortuosity, wall shear stress, resistance and associated cerebral oxygen extraction fraction and metabolism were also quantified. With the use of vasodilatory stimulus, multifaceted cerebrovascular reactivity (CVR) was characterized in vivo. By extending the classic Evans blue assay to in vivo, permeability of the blood-brain barrier (BBB) was dynamically evaluated. The utility of this enabling technique was examined by studying cerebrovascular alterations in an established mouse model of high-fat diet-induced obesity. Our results revealed increased vascular density, reduced arterial flow, enhanced oxygen extraction, impaired BBB integrity, and increased multifaceted CVR in the obese brain. Interestingly, the 'counterintuitive' increase of CVR was supported by the elevated active endothelial nitric oxide synthase in the obese mouse. Providing comprehensive and quantitative insights into cerebral microvessels and their responses under pathological conditions, this technique opens a new door to mechanistic studies of the cerebral small vessel disease and its implications in neurodegeneration and stroke.
Assuntos
Barreira Hematoencefálica/diagnóstico por imagem , Encéfalo/irrigação sanguínea , Encéfalo/diagnóstico por imagem , Microvasos/diagnóstico por imagem , Obesidade/diagnóstico por imagem , Técnicas Fotoacústicas , Animais , Modelos Animais de Doenças , Masculino , Camundongos , MicroscopiaRESUMO
Gold-silver nanocages (GSNCs) are widely used in cancer imaging and therapy due to excellent biocompatibility, internal hollow structures, and tunable optical properties. However, their possible responses toward the tumor microenvironment are still not well understood. In this study, it is demonstrated that a kind of relatively small sized (35 nm) and partially hollow GSNCs (absorbance centered at 532 nm) can enhance the intrinsic photoacoustic imaging performances for blood vessels around tumor sites. More importantly, the high concentration of glutathione around the tumor cells' microenvironment may induce the aggregation, disintegration, and agglomeration of these GSNCs sequentially, allowing significant shifts in the absorbance spectrum of GSNCs to the near-infrared (NIR) region. This enhanced absorbance in the NIR region entails the significant photothermal therapy (PTT) effect. In vivo experiments, including photoacoustic microscopy (PAM) for cancer diagnosis and PTT in tumor model mice, also show coincident consequences. Taken together, the slightly hollow GSNCs may assist PAM-based tumor diagnosis and induce a tumor targeted PTT effect. This work paves a new avenue for the development of an alternative tumor diagnostic and therapeutic strategy.
Assuntos
Glutationa/química , Ouro/química , Hipertermia Induzida , Nanoestruturas/química , Neoplasias/diagnóstico , Neoplasias/terapia , Fototerapia , Prata/química , Nanomedicina Teranóstica , Microambiente TumoralRESUMO
Choroidal neovascularization (CNV) is a major cause of vision loss that consists of abnormal growth of new blood vessels from the choroidal vasculature. High resolution in vivo imaging of animal models is essential to better elucidate and conduct research on CNV. This study evaluates a novel multimodal imaging platform combining optical coherence tomography (OCT) and photoacoustic microscopy (PAM). Using real-time OCT guidance subretinal injection to induce and multimodality imaging system to monitor CNV over time in rabbit eyes. The significance of our work lies in providing the optimal setting and conditions to make use of the OCT image guided system to improve the consistency and reproducibility of experimental results in subretinal injection induced CNV model in rabbits. For the first time, this study successfully demonstrated the dual-modality PAM-OCT system, without using exogenous contrast agents, can detect and visualize CNV in the rabbit eye with high resolution. This is promising system for diagnosing and monitoring CNV.
Assuntos
Neovascularização de Coroide/diagnóstico por imagem , Técnicas de Diagnóstico Oftalmológico , Microscopia Acústica/métodos , Tomografia de Coerência Óptica/métodos , Animais , Neovascularização de Coroide/etiologia , Injeções Intraoculares/efeitos adversos , Imagem Multimodal , Coelhos , Reprodutibilidade dos TestesRESUMO
The opacity of conventional ultrasound transducers can impede the miniaturization and workflow of current photoacoustic systems. In particular, optical-resolution photoacoustic microscopy (OR-PAM) requires the coaxial alignment of optical illumination and acoustic-detection paths through complex beam combiners and a thick coupling medium. To overcome these hurdles, we developed a novel OR-PAM method on the basis of our recently reported transparent lithium niobate (LiNbO3) ultrasound transducer (Dangi et al., Optics Letters, 2019), which was centered at 13 MHz ultrasound frequency with 60% photoacoustic bandwidth. To test the feasibility of wearable OR-PAM, optical-only raster scanning of focused light through a transducer was performed while the transducer was fixed above the imaging subject. Imaging experiments on resolution targets and carbon fibers demonstrated a lateral resolution of 8.5 µm. Further, we demonstrated vasculature mapping using chicken embryos and melanoma depth profiling using tissue phantoms. In conclusion, the proposed OR-PAM system using a low-cost transparent LiNbO3 window transducer has a promising future in wearable and high-throughput imaging applications, e.g., integration with conventional optical microscopy to enable a multimodal microscopy platform capable of ultrasound stimulation.
RESUMO
Photoacoustic microscopy (PAM) is an emerging biomedical imaging technology capable of quantitative measurement of the microvascular blood flow by correlation analysis. However, the computational cost is high, limiting its applications. Here, we report a parallel computation design based on graphics processing unit (GPU) for high-speed quantification of blood flow in PAM. Two strategies were utilized to improve the computational efficiency. First, the correlation method in the algorithm was optimized to avoid redundant computation and a parallel computing structure was designed. Second, the parallel design was realized on GPU and optimized by maximizing the utilization of computing resource in GPU. The detailed timings and speedup for each calculation step were given and the MATLAB and C/C++ code versions based on CPU were presented as a comparison. Full performance test shows that a stable speedup of ~80-fold could be achieved with the same calculation accuracy and the computation time could be reduced from minutes to just several seconds with the imaging size ranging from 1 × 1 mm2 to 2 × 2 mm2. Our design accelerates PAM-based blood flow measurement and paves the way for real-time PAM imaging and processing by significantly improving the computational efficiency.
Assuntos
Imageamento Tridimensional , Microscopia , Técnicas Fotoacústicas , Fluxo Sanguíneo Regional/fisiologia , Algoritmos , Animais , Gráficos por Computador , Masculino , CamundongosRESUMO
In this study, dual-modal fluorescence and photoacoustic microscopy was performed for noninvasive and functional in vivo imaging of inflammation induced by green fluorescent protein (GFP) transfected bacteria in mice ear. Our imaging results demonstrated that the multimodal imaging technique is able to monitor the tissue immunovascular responses to infections with molecular specificity. Our study also indicated that the combination of photoacoustic and fluorescence microscopy imaging can simultaneously track the biochemical changes including the bacterial distribution and morphological change of blood vessels in the biological tissues with high resolution and enhanced sensitivity. Consequently, the developed method paves a new avenue for improving the understanding of the pathology mechanism of inflammation.