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Multi-scale photoacoustic remote sensing (PARS) (Conference Presentation) 多尺度光声遥感(PARS)(会议报告)
Pub Date : 2016-04-27 DOI: 10.1117/12.2211467
P. Haji Reza, K. Bell, W. Shi, R. Zemp
We introduce a novel multi-scale photoacoustic remote sensing (PARS) imaging system. Our system can provide optical resolution details for superficial structures as well as acoustic resolution for deep-tissue imaging down to 5 cm, in a non-contact setting. PARS system does not require any contact with the sample or ultrasound coupling medium. The optical resolution PARS (OR-OARS) system uses optically focused pulsed excitation with optical detection of photoacoustic signatures using a long-coherence interrogation beam co-focused and co-scanned with the excitation spot. In the OR-PARS initial pressures are sampled right at their subsurface origin where acoustic pressures are largest. The Acoustic resolution PARS (AR-PARS) picks up the surface oscillation of the tissue caused by generated photoacoustic signal using a modified version of Michelson interferometry. By taking advantage of 4-meters polarization maintaining single-mode fiber and a green fiber laser we have generated a multi-wavelength source using stimulated Raman scattering. Remote functional imaging using this multi-wavelength excitation source and PARS detection mechanism has been demonstrated. The oxygen saturation estimations are shown for both phantom and in vivo studies. Images of blood vessel structures for an In vivo chicken embryo model is demonstrated. The Phantom studies indicates ~3µm and ~300µm lateral resolution for OR-PARS and AR-PARS respectively. To the best of our knowledge this is the first dual modality non-contact optical and acoustic resolution system used for in vivo imaging.
介绍了一种新型的多尺度光声遥感成像系统。我们的系统可以为表面结构提供光学分辨率细节,也可以在非接触式环境下为5厘米的深层组织成像提供声学分辨率。PARS系统不需要与样品或超声耦合介质有任何接触。光学分辨率PARS (OR-OARS)系统使用光聚焦脉冲激励和光声特征的光学检测,使用长相干询问光束与激励点共聚焦和共扫描。在OR-PARS中,初始压力是在声压最大的地下起源处采样的。声学分辨率PARS (AR-PARS)采用一种改进的迈克尔逊干涉测量法,提取由产生的光声信号引起的组织表面振荡。利用保持4米偏振的单模光纤和绿色光纤激光器,利用受激拉曼散射产生了多波长光源。利用该多波长激发源进行远程功能成像,并验证了PARS检测机制。氧饱和度估计显示了幻影和体内研究。展示了鸡体内胚胎模型的血管结构图像。Phantom研究表明,OR-PARS和AR-PARS的横向分辨率分别为~3µm和~300µm。据我们所知,这是第一个用于体内成像的双模态非接触式光学和声学分辨率系统。
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引用次数: 0
Fourier-interpolation superresolution optical fluctuation imaging (fSOFi) (Conference Presentation) 傅里叶插值超分辨率光学波动成像(fSOFi)(会议报告)
Pub Date : 2016-04-27 DOI: 10.1117/12.2212289
J. Enderlein, S. Stein, A. Huss, D. Hähnel, I. Gregor
Stochastic Optical Fluctuation Imaging (SOFI) is a superresolution fluorescence microscopy technique which allows to enhance the spatial resolution of an image by evaluating the temporal fluctuations of blinking fluorescent emitters. SOFI is not based on the identification and localization of single molecules such as in the widely used Photoactivation Localization Microsopy (PALM) or Stochastic Optical Reconstruction Microscopy (STORM), but computes a superresolved image via temporal cumulants from a recorded movie. A technical challenge hereby is that, when directly applying the SOFI algorithm to a movie of raw images, the pixel size of the final SOFI image is the same as that of the original images, which becomes problematic when the final SOFI resolution is much smaller than this value. In the past, sophisticated cross-correlation schemes have been used for tackling this problem. Here, we present an alternative, exact, straightforward, and simple solution using an interpolation scheme based on Fourier transforms. We exemplify the method on simulated and experimental data.
随机光学波动成像(SOFI)是一种超分辨率荧光显微镜技术,它可以通过评估闪烁荧光发射器的时间波动来提高图像的空间分辨率。SOFI不像广泛使用的光激活定位显微镜(PALM)或随机光学重建显微镜(STORM)那样基于单个分子的识别和定位,而是通过记录电影的时间累积量计算超分辨图像。这里的一个技术挑战是,当直接将SOFI算法应用于原始图像的电影时,最终SOFI图像的像素大小与原始图像的像素大小相同,当最终SOFI分辨率远小于此值时,就会出现问题。在过去,复杂的互相关方案被用来解决这个问题。在这里,我们提出了一种替代的,精确的,直接的,简单的解决方案,使用基于傅里叶变换的插值方案。通过仿真和实验数据对该方法进行了验证。
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引用次数: 0
Intraoperative detection and elimination of microscopic tumors in head and neck (Conference Presentation) 术中头颈部显微肿瘤的检测与消除(会议报告)
Pub Date : 2016-04-27 DOI: 10.1117/12.2217942
E. Lukianova-Hleb, Yoo-shin Kim, Ihar Belatsarkouski, E. Hanna, A. Gillenwater, B. O'Neill, D. Lapotko
Failure of cancer surgery to intraoperatively detect and eliminate microscopic residual disease (MRD) causes lethal recurrence and metastases, whereas removal of important normal tissues causes excessive morbidity. We report plasmonic nanobubble (PNB) surgical technology to intraoperatively detect and eliminate MRD in surgical bed. PNBs were generated in vivo in head and neck cancer cells by systemically targeting tumor with gold colloids and locally-applied near-infrared low energy short laser pulse, and were simultaneously detected with acoustic probe. In mouse models of head and neck squamous cell carcinoma, single cancer cells and MRD (undetectable with standard histological methods) were instantaneously non-invasively detected in solid tissue in surgical bed. In resectable MRD, PNB-guided surgery prevented local recurrence and delivered 100% tumor-free survival. In unresectable MRD, PNB nano-surgery improved survival by two-fold compared to standard surgery. PNB metrics correlated with the tumor recurrence rate. PNB surgical technology precisely detects and immediately eliminates MRD at macro- and micro-scale in a simple and safe intraoperative procedure.
癌症手术未能在术中发现并消除显微残留病变(MRD)会导致致命的复发和转移,而切除重要的正常组织则会导致过高的发病率。我们报道了等离子体纳米泡(PNB)手术技术在术中检测和消除手术床上的MRD。利用金胶体系统靶向肿瘤,局部施加近红外低能量短激光脉冲,在头颈部癌细胞体内产生PNBs,并与声探针同步检测。在头颈部鳞状细胞癌小鼠模型中,在手术床上的实体组织中,可以即时无创地检测到单个癌细胞和MRD(标准组织学方法无法检测到)。在可切除的MRD中,pnb引导的手术预防了局部复发,并提供了100%的无肿瘤生存率。在不可切除的MRD中,与标准手术相比,PNB纳米手术将生存率提高了两倍。PNB指标与肿瘤复发率相关。PNB手术技术在简单安全的术中精确检测并立即消除宏观和微观尺度的MRD。
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引用次数: 0
Detection of esophageal cancer cell by photoelectrochemical Cu2O/ZnO biosensor (Conference Presentation) 光电化学Cu2O/ZnO生物传感器检测食管癌细胞(会议报告)
Pub Date : 2016-04-27 DOI: 10.1117/12.2212073
Chao-Hsin Hsu, Chengjiang Chu, Weichung Chen, I-Chen Wu, Ming-Tsang Wu, C. Kuo, R. Tsiang, Hsiang-Chen Wang
We have demonstrated a Cu2O/ZnO nanorods (NRs) array p-n heterostructures photoelectrochemical biosensor. The electrodeposition of Cu2O at pH 12 acquired the preferably (111) lattice planes, resulting in the largest interfacial electric field between Cu2O and ZnO, which finally led to the highest separation efficiency of photogenerated charge carriers. High verticality ZnO nanorods by seed layer and thermal annealing assist the hydrothermal growth. The optimized Cu2O/ZnO NRs array p-n heterostructures exhibited enhanced PEC performance, such as elevated photocurrent and photoconversion efficiency, as well as excellent sensing performance for the sensitive detection of four strains of different races and different degree of cancer cell which made the device self-powered. We got spectral response characteristics and operating wavelength range of biosensor, and to verify the biological characteristics of cancer cells wafer react with different stages of cancer characterized by a cancer measured reaction experiment.
我们展示了一种Cu2O/ZnO纳米棒(NRs)阵列p-n异质结构光电化学生物传感器。在pH值为12的条件下电沉积Cu2O获得了较好的(111)晶格面,使得Cu2O与ZnO之间的界面电场最大,最终使得光生载流子的分离效率最高。通过种子层和热退火制备的高垂直度ZnO纳米棒有助于水热生长。优化后的Cu2O/ZnO NRs阵列p-n异质结构具有更高的光电电流和光转换效率,对不同种族的四种菌株和不同程度的癌细胞具有良好的传感性能,使器件具有自供电能力。我们得到了生物传感器的光谱响应特性和工作波长范围,并通过癌症测量反应实验验证了癌细胞与不同癌症阶段反应的生物学特性。
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引用次数: 0
Investigate the variation in optical redox ratio of epicardial adipose tissue in patients with CAD through auto-fluorescence metabolic molecular image (Conference Presentation) 应用自体荧光代谢分子图像研究冠心病患者心外膜脂肪组织光学氧化还原比的变化(会议报告)
Pub Date : 2016-04-27 DOI: 10.1117/12.2212606
Lun-Zhang Guo, Tzung-Dau Wang, Jong-Wei Lin, Tzu-Ming Liu
In recent years, it has been suggested that epicardial adipose tissue (EAT) plays an important role in development of coronary artery disease (CAD) and diabetes mellitus (DM). In this article, we used two-photon fluoresce microscope to measure the fluorescence metabolic image of EAT, which obtained from the patient with/without CAD/DM. We used 740nm and 890nm infrared light to excite the auto-fluorescence of metabolic molecules NADH and FAD respectively. We collected the fluorescence signal at wavelength 450nm to 500nm and 500nm to 550nm to obtain the metabolic image. Through the image, we computed the redox ratio (NADH/FAD) by analyzing the intensity. The preliminary result showed that the redox ratio increase in the patients with CAD. It indicates EAT adipocytes of patient with CAD have decreased cellular metabolic activity. But there were no significant variation of redox ratio in the patients with DM.
近年来,心外膜脂肪组织(EAT)在冠状动脉疾病(CAD)和糖尿病(DM)的发生发展中起着重要作用。在本文中,我们使用双光子荧光显微镜测量了CAD/DM患者/非CAD/DM患者的EAT荧光代谢图像。我们分别用740nm和890nm红外光激发代谢分子NADH和FAD的自身荧光。我们采集波长为450nm ~ 500nm和500nm ~ 550nm的荧光信号,得到代谢图像。通过图像分析强度,计算出氧化还原比(NADH/FAD)。初步结果表明,冠心病患者的氧化还原比升高。提示冠心病患者的EAT脂肪细胞代谢活性降低。而DM患者的氧化还原比无明显变化。
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引用次数: 0
Screening photoswitching properties of synthesized BODIPY-based fluorophores for multispectral superresolution microscopy (MSSRM) (Conference Presentation) 用于多光谱超分辨显微镜(MSSRM)的合成基于bodipy的荧光团的光开关特性筛选(会议报告)
Pub Date : 2016-04-27 DOI: 10.1117/12.2213462
Amy M. Bittel, Isaac S. Saldivar, Xiaolin Nan, Summer L. Gibbs
Single-molecule localization microscopy (SMLM) utilizes photoswitchable fluorophores to detect biological entities with 10-20 nm resolution. Multispectral superresolution microscopy (MSSRM) extends SMLM functionality by improving its spectral resolution up to 5 fold facilitating imaging of multicomponent cellular structures or signaling pathways. Current commercial fluorophores are not ideal for MSSRM as they are not designed to photoswitch and do not adequately cover the visible and far-red spectral regions required for MSSRM imaging. To obtain optimal MSSRM spatial and spectral resolution, fluorophores with narrow emission spectra and controllable photoswitching properties are necessary. Herein, a library of BODIPY-based fluorophores was synthesized and characterized to create optimal photoswitchable fluorophores for MSSRM. BODIPY was chosen as the core structure as it is photostable, has high quantum yield, and controllable photoswitching. The BODIPY core was modified through the addition of various aromatic moieties, resulting in a spectrally diverse library. Photoswitching properties were characterized using a novel polyvinyl alcohol (PVA) based film methodology to isolate single molecules. The PVA film methodology enabled photoswitching assessment without the need for protein conjugation, greatly improving screening efficiency of the BODIPY library. Additionally, image buffer conditions were optimized for the BODIPY-based fluorophores through systematic testing of oxygen scavenger systems, redox components, and additives. Through screening the photoswitching properties of BODIPY-based compounds in PVA films with optimized imaging buffer we identified novel fluorophores well suited for SMLM and MSSRM.
单分子定位显微镜(SMLM)利用光开关荧光团检测生物实体,分辨率为10-20纳米。多光谱超分辨率显微镜(MSSRM)通过将其光谱分辨率提高到5倍,扩展了SMLM功能,促进了多组分细胞结构或信号通路的成像。目前的商用荧光团不适合MSSRM,因为它们没有设计成光开关,也没有充分覆盖MSSRM成像所需的可见光和远红色光谱区域。为了获得最佳的MSSRM空间和光谱分辨率,需要具有窄发射光谱和可控光电开关特性的荧光团。本文合成了一个基于bodipy的荧光团库,并对其进行了表征,以创建用于MSSRM的最佳光切换荧光团。之所以选择BODIPY作为核心结构,是因为它具有光稳定、高量子产率和可控的光开关特性。通过添加各种芳香基团对BODIPY核心进行了修改,从而形成了一个光谱多样化的库。利用新型聚乙烯醇(PVA)薄膜分离方法表征了单分子的光开关特性。PVA薄膜方法使光开关评估无需蛋白质偶联,大大提高了BODIPY文库的筛选效率。此外,通过对氧清除系统、氧化还原组分和添加剂的系统测试,优化了基于bodipy的荧光团的图像缓冲条件。通过筛选基于bodipi的化合物在PVA薄膜中的光电开关特性,并优化成像缓冲液,我们确定了适合于SMLM和MSSRM的新型荧光团。
{"title":"Screening photoswitching properties of synthesized BODIPY-based fluorophores for multispectral superresolution microscopy (MSSRM) (Conference Presentation)","authors":"Amy M. Bittel, Isaac S. Saldivar, Xiaolin Nan, Summer L. Gibbs","doi":"10.1117/12.2213462","DOIUrl":"https://doi.org/10.1117/12.2213462","url":null,"abstract":"Single-molecule localization microscopy (SMLM) utilizes photoswitchable fluorophores to detect biological entities with 10-20 nm resolution. Multispectral superresolution microscopy (MSSRM) extends SMLM functionality by improving its spectral resolution up to 5 fold facilitating imaging of multicomponent cellular structures or signaling pathways. Current commercial fluorophores are not ideal for MSSRM as they are not designed to photoswitch and do not adequately cover the visible and far-red spectral regions required for MSSRM imaging. To obtain optimal MSSRM spatial and spectral resolution, fluorophores with narrow emission spectra and controllable photoswitching properties are necessary. Herein, a library of BODIPY-based fluorophores was synthesized and characterized to create optimal photoswitchable fluorophores for MSSRM. BODIPY was chosen as the core structure as it is photostable, has high quantum yield, and controllable photoswitching. The BODIPY core was modified through the addition of various aromatic moieties, resulting in a spectrally diverse library. Photoswitching properties were characterized using a novel polyvinyl alcohol (PVA) based film methodology to isolate single molecules. The PVA film methodology enabled photoswitching assessment without the need for protein conjugation, greatly improving screening efficiency of the BODIPY library. Additionally, image buffer conditions were optimized for the BODIPY-based fluorophores through systematic testing of oxygen scavenger systems, redox components, and additives. Through screening the photoswitching properties of BODIPY-based compounds in PVA films with optimized imaging buffer we identified novel fluorophores well suited for SMLM and MSSRM.","PeriodicalId":227483,"journal":{"name":"SPIE BiOS","volume":"8 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"120903805","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Use of a highly transparent zebrafish mutant for investigations in the development of the vertebrate auditory system (Conference Presentation) 利用高度透明的斑马鱼突变体研究脊椎动物听觉系统的发育(会议报告)
Pub Date : 2016-04-27 DOI: 10.1117/12.2213087
Anna M. Wisniowiecki, Scott P. Mattison, Sangmin Kim, B. Riley, B. Applegate
Zebrafish, an auditory specialist among fish, offer analogous auditory structures to vertebrates and is a model for hearing and deafness in vertebrates, including humans. Nevertheless, many questions remain on the basic mechanics of the auditory pathway. Phase-sensitive Optical Coherence Tomography has been proven as valuable technique for functional vibrometric measurements in the murine ear. Such measurements are key to building a complete understanding of auditory mechanics. The application of such techniques in the zebrafish is impeded by the high level of pigmentation, which develops superior to the transverse plane and envelops the auditory system superficially. A zebrafish double mutant for nacre and roy (mitfa-/- ;roya-/- [casper]), which exhibits defects for neural-crest derived melanocytes and iridophores, at all stages of development, is pursued to improve image quality and sensitivity for functional imaging. So far our investigations with the casper mutants have enabled the identification of the specialized hearing organs, fluid-filled canal connecting the ears, and sub-structures of the semicircular canals. In our previous work with wild-type zebrafish, we were only able to identify and observe stimulated vibration of the largest structures, specifically the anterior swim bladder and tripus ossicle, even among small, larval specimen, with fully developed inner ears. In conclusion, this genetic mutant will enable the study of the dynamics of the zebrafish ear from the early larval stages all the way into adulthood.
斑马鱼是鱼类中的听觉专家,具有与脊椎动物相似的听觉结构,是包括人类在内的脊椎动物听力和耳聋的模型。然而,关于听觉通路的基本机制仍然存在许多问题。相敏光学相干层析成像已被证明是一种有价值的功能振动测量技术。这些测量是建立对听觉机制完整理解的关键。这种技术在斑马鱼中的应用受到高度色素沉着的阻碍,这些色素沉着发展到横向平面之上,并表面包裹着听觉系统。一种斑马鱼珠质和黄酮双突变体(mitfa-/-;roya-/- [casper]),在发育的所有阶段都表现出神经嵴来源的黑素细胞和虹膜细胞的缺陷,旨在提高功能成像的图像质量和灵敏度。到目前为止,我们对卡斯马突变体的研究已经能够识别专门的听觉器官,连接耳朵的充满液体的管道和半圆形管道的亚结构。在我们之前对野生型斑马鱼的研究中,我们只能识别和观察到最大结构的刺激振动,特别是前鱼鳔和三听骨,甚至在内耳发育完全的小型幼虫标本中也是如此。总之,这种基因突变将使斑马鱼耳朵从早期幼虫阶段一直到成年的动力学研究成为可能。
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引用次数: 1
In-vivo continuous monitoring of mixed venous oxygen saturation by photoacoustic transesophageal echocardiography (Conference Presentation) 经食管光声超声心动图在体内连续监测混合静脉氧饱和度(会议报告)
Pub Date : 2016-04-27 DOI: 10.1117/12.2211112
Li Li, B. Subramaniam, A. Aguirre, Michael N. Andrawes, G. Tearney
Mixed venous oxygen saturation (SvO2), measured from pulmonary arteries, is a gold-standard measure of the dynamic balance between the oxygen supply and demand in the body. In critical care, continuous monitoring of SvO2 plays a vital role in early detection of circulatory shock and guiding goal-oriented resuscitation. In current clinical practice, SvO2 is measured by invasive pulmonary artery catheters (PAC), which are associated with a 10% risk of severe complications. To address the unmet clinical need for a non-invasive SvO2 monitor, we are developing a new technology termed photoacoustic transesophageal echocardiography (PA-TEE). PA-TEE integrates transesophageal echocardiography with photoacoustic oximetry, and enables continuous assessment of SvO2 through an esophageal probe that can be inserted into the body in a minimally invasive manner. We have constructed a clinically translatable PA-TEE prototype, which features a mobile OPO laser, a modified ultrasonography console and a dual-modality esophageal probe. Comprised of a rotatable acoustic array detector, a flexible optical fiber bundle and a light-integrating acoustic lens, the oximetric probe has an outer diameter smaller than 15 mm and will be tolerable for most patients. Through custom-made C++/Qt software, our device acquires and displays ultrasonic and photoacoustic images in real time to guide the deployment of the probe. SvO2 is calculated on-line and updated every second. PA-TEE has now been used to evaluate SvO2 in living swine. Our findings show that changing the fraction of oxygen in the inspired gas modulates SvO2 measured by PA-TEE. Statistic comparison between SvO2 measurements from PA-TEE in vivo the gold-standard laboratorial analysis on blood samples drawn from PACs will be presented.
混合静脉氧饱和度(SvO2),由肺动脉测量,是衡量体内氧供需动态平衡的金标准。在重症监护中,持续监测SvO2对早期发现循环休克和指导目标导向的复苏具有重要作用。在目前的临床实践中,SvO2是通过有创肺动脉导管(PAC)测量的,这与10%的严重并发症风险相关。为了满足临床对无创SvO2监测的需求,我们正在开发一种新技术,称为光声经食管超声心动图(PA-TEE)。PA-TEE将经食管超声心动图与光声血氧仪相结合,通过可插入体内的食管探头,以微创方式连续评估SvO2。我们构建了一个临床可翻译的PA-TEE原型,它具有一个移动的OPO激光器,一个改进的超声检查控制台和一个双模态食管探头。由一个可旋转的声阵列探测器、一个柔性光纤束和一个光集成声透镜组成,血氧测量探头的外径小于15毫米,对大多数患者来说是可以忍受的。我们的设备通过定制的c++ /Qt软件,实时采集并显示超声波和光声图像,指导探头的部署。SvO2是在线计算并每秒更新的。PA-TEE现已用于活猪体内SvO2的评价。我们的研究结果表明,改变吸入气体中氧气的比例可以调节PA-TEE测量的SvO2。将介绍PA-TEE在体内测量SvO2的统计比较,以及从PACs抽取的血液样本的金标准实验室分析。
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引用次数: 2
All-optical pulse-echo ultrasound probe for intravascular imaging (Conference Presentation) 用于血管内成像的全光脉冲回波超声探头(会议报告)
Pub Date : 2016-04-27 DOI: 10.1117/12.2213757
R. Colchester, S. Noimark, C. Mosse, E. Zhang, P. Beard, I. Parkin, I. Papakonstantinou, A. Desjardins
High frequency ultrasound probes such as intravascular ultrasound (IVUS) and intracardiac echocardiography (ICE) catheters can be invaluable for guiding minimally invasive medical procedures in cardiology such as coronary stent placement and ablation. With current-generation ultrasound probes, ultrasound is generated and received electrically. The complexities involved with fabricating these electrical probes can result in high costs that limit their clinical applicability. Additionally, it can be challenging to achieve wide transmission bandwidths and adequate wideband reception sensitivity with small piezoelectric elements. Optical methods for transmitting and receiving ultrasound are emerging as alternatives to their electrical counterparts. They offer several distinguishing advantages, including the potential to generate and detect the broadband ultrasound fields (tens of MHz) required for high resolution imaging. In this study, we developed a miniature, side-looking, pulse-echo ultrasound probe for intravascular imaging, with fibre-optic transmission and reception. The axial resolution was better than 70 microns, and the imaging depth in tissue was greater than 1 cm. Ultrasound transmission was performed by photoacoustic excitation of a carbon nanotube/polydimethylsiloxane composite material; ultrasound reception, with a fibre-optic Fabry-Perot cavity. Ex vivo tissue studies, which included healthy swine tissue and diseased human tissue, demonstrated the strong potential of this technique. To our knowledge, this is the first study to achieve an all-optical pulse-echo ultrasound probe for intravascular imaging. The potential for performing all-optical B-mode imaging (2D and 3D) with virtual arrays of transmit/receive elements, and hybrid imaging with pulse-echo ultrasound and photoacoustic sensing are discussed.
高频超声探头,如血管内超声(IVUS)和心内超声心动图(ICE)导管,对于指导心脏病学中的微创医疗程序,如冠状动脉支架置入术和消融术,是非常宝贵的。使用当前一代超声波探头,超声波是用电产生和接收的。制造这些电探针的复杂性可能导致高成本,限制了它们的临床应用。此外,利用小型压电元件实现较宽的传输带宽和足够的宽带接收灵敏度可能具有挑战性。传输和接收超声波的光学方法正在成为其电子对应物的替代品。它们有几个显著的优势,包括产生和检测高分辨率成像所需的宽带超声场(数十兆赫)的潜力。在这项研究中,我们开发了一种微型、侧视、脉冲回波超声探头,用于血管内成像,具有光纤传输和接收。轴向分辨率优于70微米,组织成像深度大于1 cm。采用光声激发碳纳米管/聚二甲基硅氧烷复合材料实现超声传输;超声波接收,光纤法布里-珀罗腔。包括健康猪组织和患病人组织在内的离体组织研究证明了该技术的强大潜力。据我们所知,这是第一个实现血管内成像全光脉冲回波超声探头的研究。讨论了利用发射/接收元件的虚拟阵列进行全光b模成像(2D和3D)以及脉冲回波超声和光声传感混合成像的潜力。
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引用次数: 1
Advanced multiphoton methods for in vitro and in vivo functional imaging of mouse retinal neurons (Conference Presentation) 小鼠视网膜神经元体外和体内功能成像的先进多光子方法(会议报告)
Pub Date : 2016-04-27 DOI: 10.1117/12.2213185
N. Cohen, Adi Schejter, N. Farah, S. Shoham
Studying the responses of retinal ganglion cell (RGC) populations has major significance in vision research. Multiphoton imaging of optogenetic probes has recently become the leading approach for visualizing neural populations and has specific advantages for imaging retinal activity during visual stimulation, because it leads to reduced direct photoreceptor excitation. However, multiphoton retinal activity imaging is not straightforward: point-by-point scanning leads to repeated neural excitation while optical access through the rodent eye in vivo has proven highly challenging. Here, we present two enabling optical designs for multiphoton imaging of responses to visual stimuli in mouse retinas expressing calcium indicators. First, we present an imaging solution based on Scanning Line Temporal Focusing (SLITE) for rapidly imaging neuronal activity in vitro. In this design, we scan a temporally focused line rather than a point, increasing the scan speed and reducing the impact of repeated excitation, while maintaining high optical sectioning. Second, we present the first in vivo demonstration of two-photon imaging of RGC activity in the mouse retina. To obtain these cellular resolution recordings we integrated an illumination path into a correction-free imaging system designed using an optical model of the mouse eye. This system can image at multiple depths using an electronically tunable lens integrated into its optical path. The new optical designs presented here overcome a number of outstanding obstacles, allowing the study of rapid calcium- and potentially even voltage-indicator signals both in vitro and in vivo, thereby bringing us a step closer toward distributed monitoring of action potentials.
研究视网膜神经节细胞(RGC)群体的反应在视觉研究中具有重要意义。光遗传探针的多光子成像最近成为可视化神经群的主要方法,并且在视觉刺激期间成像视网膜活动具有特定的优势,因为它导致直接光感受器激发减少。然而,多光子视网膜活动成像并不简单:逐点扫描导致重复的神经兴奋,而通过啮齿动物眼睛的光学访问在体内已被证明是极具挑战性的。在这里,我们提出了两种可行的光学设计,用于表达钙指标的小鼠视网膜对视觉刺激的反应的多光子成像。首先,我们提出了一种基于扫描线时间聚焦(SLITE)的成像解决方案,用于快速成像体外神经元活动。在本设计中,我们扫描一个暂时聚焦的线而不是一个点,提高了扫描速度,减少了重复激发的影响,同时保持了高光学切片。其次,我们首次在小鼠视网膜上展示了RGC活动的双光子成像。为了获得这些细胞分辨率记录,我们将照明路径集成到使用小鼠眼睛光学模型设计的免校正成像系统中。该系统可以使用集成在其光路中的电子可调透镜在多个深度成像。这里提出的新的光学设计克服了许多突出的障碍,允许在体外和体内快速研究钙甚至潜在的电压指示信号,从而使我们更接近动作电位的分布式监测。
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引用次数: 0
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