光声结构与功能成像技术研究进展

光声成像技术利用短脉冲激光激发产生光声信号,可重建出组织的光吸收分布图像,它结合了纯光学成像的高对比度和纯声学成像的高分辨率特性.光声成像技术不仅能够有效的刻画生物组织结构,还能够精确实现无损功能成像,为研究生物组织的形态结构,生理、病理特征,代谢功能等提供了全新手段.本文简要分析了光声信号产生的机理,总结报道了目前实验室几套典型的成像系统及其最新应用进展,指出光声成像作为一种新型的生物医学成像方法,可望引发生物医学影像领域的一次革新.     

窄等离子体吸收谱线宽度的纳米金锥用于光声成像

无损光声成像技术结合了纯光学成像高选择特性和纯超声成像中深穿透特性的优点,克服了光散射限制,实现了对活体深层组织的高分辨、高对比度成像。该成像技术对内源物质例如脱氧血红蛋白、含氧血红蛋白、黑色素、脂质等进行成像,提供了活体生物组织结构和功能信息,已经在生物医学领域表现出巨大的应用前景。然而,很多与病理过程相关的特征分子的光吸收能力较弱,在活体环境中难以被光声成像系统所识别,从而限制了光声成像技术的应用范围。基于功能纳米探针的光声成像-光声分子成像极大拓展光声成像的应用范围,可以在活体层面对病理过程进行分子水平的定性和定量研究,将为实现目标疾病的早期诊断提供强大的技术支持。本文发展在近红外具有窄吸收线宽(半高宽仅为60 nm)的纳米金锥作为新型的光声探针。通过选择不同径长比的纳米金锥,可以任意调节纳米金锥的吸收峰。通过调谐激光器的波长,可实现对不同吸收峰纳米金锥的选择性激发。纳米金锥将有可能用于多光谱光声成像,实现对不同靶标的目标分子探测。     

基于压缩感知理论的光声成像方法研究现状

光声成像是一种新兴的无损生物医学成像方法,因其兼具高灵敏的光学对比度和超声能够对深层组织进行高分辨成像的优点,已经成为当前生物医学成像领域发展最快的技术之一。光声成像的光吸收对比度能够反映生物组织微小的组织病变,与血氧饱和度等多种功能和生理信息紧密相关,目前已被证明在肿瘤血管新生研究、早期癌症检测和心血管疾病诊断等方面有很大的应用潜力。基于超声阵列探测的常规光声计算层析成像系统,数据采集量大,由此导致的较低数据采集和成像速度成为制约该技术临床应用和转化的重要因素。压缩感知理论可以在远低于Nyquist采样定理的欠采样方式下,高质量重建信号,已被广泛用于信号处理和传统的医学图像重建领域。自2009年压缩感知理论被应用于光声成像以来,已有的研究结果表明,该方法为解决目前大区域光声成像的数据采集和成像速度问题提供了一条有效的途径。本文将重点介绍压缩感知理论用于光声成像的基本原理、研究现状、面临的问题和应用前景。     

基于光谱编码的血管内光声组分成像

光声成像突破了传统的光学成像和超声成像在生物组织成像领域的困境,该技术基于光声(Photoacoustic,PA)效应,脉冲激光激励下的生物组织产生超声信号,超声信号被接收后,通过反投影算法将其携带的时间信息和强度信息转化为能够反映生物组织吸收结构和分布的可视化图像。基于不同生物组织的光吸收差异,当激发光强度均匀且稳定时,光声成像反映的就是该物质对于该波长光的吸收特性。本文中,我们基于导管式的血管内光声断层扫描平台结合多波长激发的光声成像算法开发了基于光谱编码的血管内光声组分成像系统,实现了在离体血管斑块中脂质组分的定量成像,高分辨获得了脂质核心的大小形态和边界信息,表征了斑块内的脂质相对含量。     

光声成像造影剂的研究进展

光声成像(PAT)是利用光声效应获得生物组织或材料的断层图像或三维立体图像的一种成像方法,它兼具光学和声学成像的优点,从而成为目前比较有应用前景的一种成像模式。光声成像造影剂是光声成像的对比增强剂,它通过改变局部组织的声学和光学特性,提高成像对比度和分辨率,从而显著增强光声成像的成像效果,成为当前生物医学领域研究的一个热点。目前常见的光声成像造影剂主要有金纳米材料,碳纳米材料,染料相关纳米材料以及其他纳米材料,这些材料有它们独特的优势,它们尺寸小,稳定性好,具有良好的生物相容性,但在临床应用时本身又存在一些问题。本文综述了光声成像造影剂的种类并简要概述了其研究进展,并对其未来在生物医学领域的应用前景做了进一步展望。     

基于直线电机的大视场快速光声显微成像系统

光声成像技术是近年来发展的一种新型的无损医学成像技术,它是以脉冲激光作为激发源,以检测的声信号为信息载体,通过相应的图像重建算法重建组织内部结构和功能信息的成像方法。该方法结合了光学成像和声学成像的特点,可提供深层组织高分辨率和高对比度的组织层析图像,在生物医学临床诊断以及在体成像领域具有广泛的应用前景。目前光声成像的扫描方式主要有基于步进电机扫描方式和基于振镜的扫描方式,本文针对目前步进电机扫描速度慢(10 mm×10 mm;0.001帧/s),振镜扫描范围小(1 mm2)的不足,发展了基于直线电机扫描的大视场快速光声显微成像系统。同一条扫描线过程中直线电机速度最高可达200 mm/s。该技术采用逐线采集光声信号的方式,比逐点采集光声信号的步进电机快800倍。该系统对10 mm×10 mm全场扫描的扫描速度为0.8帧/s。最大可扫描视场范围可以达到50 mm×50 mm。大视场快速光声显微成像系统的发展将为生物医学提供新的成像工具。     

热声成像技术及其在生物医学中的研究进展

摘要：成像技术在疾病的诊断、治疗和监测中起着重要的作用。热声成像作为一种非电离和非侵入性的新型生物医学成像技术,结合了微波成像高对比度和超声成像高分辨率的优点。因其具有利用内源性对比剂（如水和离子）或多种外源性对比剂（或两者兼有）提供结构、功能、和分子信息的能力,在预临床和临床应用中显示出了巨大的潜力。近几十年来,由于微波辐射源和超声硬件的不断发展,热声成像技术已被广泛用于生物医学成像领域。本文阐述了热声成像的基本原理及成像特点,介绍了近年来热声成像技术在生物医学上的应用、当前在解决相应临床问题应用中的优势及研究现状,最后针对热声成像技术在现有生物医学中面临的挑战对该技术进行了展望。     

编者按

生物光子学是近几年来迅速崛起的一门生物物理学新技术, 它通过生物光子与物质的相互作用, 以图像的方式提供生物组织、细胞、分子的结构和功能信息。在医学无损伤诊断、细胞内分子过程的探测和脑功能成像方面,展示了良好的应用前景。本期刊登的王进军、王毅和屈军乐等的三篇论文反映了生物光子学研究在我国的兴起。王进军等应用荧光共振能量转移（简称FRET）研究了活细胞内蛋白激酶RKA活性的时空变化; 屈军乐等介绍了一种新型光学相干层析成像技术,并用它得到了视网膜上单个锥状细胞的图像; 王毅等则报道了一种新的研究生物组织结构的光声成像方法。我们希望,这三篇论文的发表能激起读者对生物光子成像技术的兴趣,并使这一技术在生命科学研究中受到更多的 关注。     

基于非线性热扩散效应的亚衍射极限光声二次谐波显微成像技术

本文提出了一种基于非线性热扩散效应的光声二次谐波显微SH-PAM成像技术,用于实现亚衍射极限光声成像。生物组织受到强度调制的高斯激光束辐射时,组织吸收光子形成高斯分布的温度场,由于热扩散系数非线性热效应引起的非线性光声PA效应,从而产生光声二次谐波信号。模拟和试验结果均表明,重建后的光声二次谐波成像的横向分辨率超过了传统光学成像分辨率。本文通过仿体样品验证了该方法的可行性,并且对人表层皮肤细胞进行了成像,以证明其对生物样品的成像能力。该方法扩展了传统光声成像的范围,为超分辨成像开辟了新的可能性,为生物医学成像和材料检测提供了新的方法。     

生物组织光声粘弹显微成像

提出一种反演生物组织粘弹信息的新型无损光声粘弹显微成像方法,它是以强度调制激光作为激发源,通过检测光声（Photoacoustic,PA）信号的相位重建组织粘弹特性分布的成像方法.实验利用不同浓度的琼脂样品来验证光声粘弹显微测量中相位随浓度变化的依赖关系.利用埋有头发丝的琼脂样品来测试这种显微方法的成像分辨率.利用具有不同粘弹性的离体生物组织来验证系统的成像能力.实验结果表明,这种新方法能够高分辨率和高对比度地重建出具有不同粘弹性的生物组织的光声粘弹显微图像,有望实现组织结晶类病变水平的显微在体检测.     

Multi-scale molecular photoacoustic tomography of gene expression

Photoacoustic tomography (PAT) is a molecular imaging technology. Unlike conventional reporter gene imaging, which is usually based on fluorescence, photoacoustic reporter gene imaging relies only on optical absorption. This work demonstrates several key merits of PAT using lacZ, one of the most widely used reporter genes in biology. We show that the expression of lacZ can be imaged by PAT as deep as 5.0 cm in biological tissue, with resolutions of ～1.0 mm and ～0.4 mm in the lateral and axial directions, respectively. We further demonstrate non-invasive, simultaneous imaging of a lacZ-expressing tumor and its surrounding microvasculature in vivo by dual-wavelength acoustic-resolution photoacoustic microscopy (AR-PAM), with a lateral resolution of 45 μm and an axial resolution of 15 μm. Finally, using optical-resolution photoacoustic microscopy (OR-PAM), we show intra-cellular localization of lacZ expression, with a lateral resolution of a fraction of a micron. These results suggest that PAT is a complementary tool to conventional optical fluorescence imaging of reporter genes for linking biological studies from the microscopic to the macroscopic scales.     

Noninvasive laser-induced photoacoustic tomography for structural and functional in vivo imaging of the brain

Imaging techniques based on optical contrast analysis can be used to visualize dynamic and functional properties of the nervous system via optical signals resulting from changes in blood volume, oxygen consumption and cellular swelling associated with brain physiology and pathology. Here we report in vivo noninvasive transdermal and transcranial imaging of the structure and function of rat brains by means of laser-induced photoacoustic tomography (PAT). The advantage of PAT over pure optical imaging is that it retains intrinsic optical contrast characteristics while taking advantage of the diffraction-limited high spatial resolution of ultrasound. We accurately mapped rat brain structures, with and without lesions, and functional cerebral hemodynamic changes in cortical blood vessels around the whisker-barrel cortex in response to whisker stimulation. We also imaged hyperoxia- and hypoxia-induced cerebral hemodynamic changes. This neuroimaging modality holds promise for applications in neurophysiology, neuropathology and neurotherapy.     

1064 nm acoustic resolution photoacoustic microscopy

Photoacoustic imaging is a noninvasive imaging technique having the advantages of high‐optical contrast and good acoustic resolution at improved imaging depths. Light transport in biological tissues is mainly characterized by strong optical scattering and absorption. Photoacoustic microscopy is capable of achieving high‐resolution images at greater depth compared to conventional optical microscopy methods. In this work, we have developed a high‐resolution, acoustic resolution photoacoustic microscopy (AR‐PAM) system in the near infra‐red (NIR) window II (NIR‐II, eg, 1064 nm) for deep tissue imaging. Higher imaging depth is achieved as the tissue scattering at 1064 nm is lesser compared to visible or near infrared window‐I (NIR‐I). Our developed system can provide a lateral resolution of 130 μm, axial resolution of 57 μm, and image up to 11 mm deep in biological tissues. This 1064‐AR‐PAM system was used for imaging sentinel lymph node and the lymph vessel in rat. Urinary bladder of rat filled with black ink was also imaged to validate the feasibility of the developed system to study deeply seated organs.      

Integrating photoacoustic microscopy with other imaging technologies for multimodal imaging

As a hybrid optical microscopic imaging technology, photoacoustic microscopy images the optical absorption contrasts and takes advantage of low acoustic scattering of biological tissues to achieve high-resolution anatomical and functional imaging. When combined with other imaging modalities, photoacoustic microscopy-based multimodal technologies can provide complementary contrast mechanisms to reveal complementary information of biological tissues. To achieve intrinsically and precisely registered images in a multimodal photoacoustic microscopy imaging system, either the ultrasonic transducer or the light source can be shared among the different imaging modalities. These technologies are the major focus of this minireview. It also covered the progress of the recently developed penta-modal photoacoustic microscopy imaging system featuring a novel dynamic focusing technique enabled by OCT contour scan.     

基于样品及点源光声信号逆卷积的光声成像方法

光声成像是一种新的生物组织成像方法,在目前的光声成像中,都是通过样品光声信号和超声探测器的脉冲响应来计算样品光吸收的投影,但是由于无法获得超声探测器较准确的脉冲响应,影响重建图像质量。提出一种新的计算样品光吸收投影的方法,从理论上给出了样品光吸收投影和样品及点源光声信号的关系,由样品及点源光声信号的逆卷积可直接计算样品光吸收的投影,点源光声信号通过聚焦入射激光直接测得。试验结果显示,重建图像和样品的相对位置、形状及尺寸完全吻合,成像系统空间分辨率达到0．3mm,证明这是一种有效的光声成像方法。     

│ω│滤波器对光声成像的影响

光声成像结合了组织纯光学成像和组织纯声学成像的优点,是一种很有潜力的无损伤的医学成像技术。本文研究了四种不同的ω滤波器,即RL滤波器,SL滤波器,改进的SL滤波器和Kwoh-R eed滤波器,利用滤波反投影算法分析了它们对光声图像重建质量的影响,由仿真和实验结果表明,Kwoh-R eed滤波器对强噪音有着很好的抑制作用,能明显的提高图像的对比度。实验所用的光源为YAG激光器,波长为532 nm,重复频率为30 H z,脉宽为7 ns,探测器为针状的PVDF膜水听器,接收面积的直径为1 mm。     

Integrated Photoacoustic Ophthalmoscopy and Spectral-domain Optical Coherence Tomography

Both the clinical diagnosis and fundamental investigation of major ocular diseases greatly benefit from various non-invasive ophthalmic imaging technologies. Existing retinal imaging modalities, such as fundus photography¹, confocal scanning laser ophthalmoscopy (cSLO)², and optical coherence tomography (OCT)³, have significant contributions in monitoring disease onsets and progressions, and developing new therapeutic strategies. However, they predominantly rely on the back-reflected photons from the retina. As a consequence, the optical absorption properties of the retina, which are usually strongly associated with retinal pathophysiology status, are inaccessible by the traditional imaging technologies.Photoacoustic ophthalmoscopy (PAOM) is an emerging retinal imaging modality that permits the detection of the optical absorption contrasts in the eye with a high sensitivity^4-7 . In PAOM nanosecond laser pulses are delivered through the pupil and scanned across the posterior eye to induce photoacoustic (PA) signals, which are detected by an unfocused ultrasonic transducer attached to the eyelid. Because of the strong optical absorption of hemoglobin and melanin, PAOM is capable of non-invasively imaging the retinal and choroidal vasculatures, and the retinal pigment epithelium (RPE) melanin at high contrasts ^6,7. More importantly, based on the well-developed spectroscopic photoacoustic imaging^5,8 , PAOM has the potential to map the hemoglobin oxygen saturation in retinal vessels, which can be critical in studying the physiology and pathology of several blinding diseases ⁹ such as diabetic retinopathy and neovascular age-related macular degeneration.Moreover, being the only existing optical-absorption-based ophthalmic imaging modality, PAOM can be integrated with well-established clinical ophthalmic imaging techniques to achieve more comprehensive anatomic and functional evaluations of the eye based on multiple optical contrasts^6,10 . In this work, we integrate PAOM and spectral-domain OCT (SD-OCT) for simultaneously in vivo retinal imaging of rat, where both optical absorption and scattering properties of the retina are revealed. The system configuration, system alignment and imaging acquisition are presented.