Name: three-dimensional ultrasonic needle tip tracking with a fiber-optic ultrasound receiver
Uploaded: 2018-08-21T13:30:00.000+00:00
Duration: 4 min 33 s
Description: Watch this Scientific Journal Video about Three-Dimensional Ultrasonic Needle Tip Tracking with a Fiber-Optic Ultrasound Receiver at JoVE.com

这项工作得到了威康信托基金的创新工程的支持 (no。WT101957) 和工程和物理科学研究理事会 (EPSRC) (no。NS/A000027/1), 由一个威康/EPSRC 中心奖 [203145 Z/16/Z & NS/A000050/1], 由欧洲研究理事会 (批准号) 的启动赠款。ERC-2012-StG, 提案 310970 MOPHIM), 并由 EPSRC 第一批 (no。EP/J010952/1)。A.L.D. 得到了伦敦大学/UCLH NIHR 综合生物医学研究中心的支持。作者感谢皇家兽医学院的工作人员在活体实验中的宝贵帮助。

1. 系统硬件
<ol><li>临床自定义超声成像探头<ol>
<li>为自定义探测器中包含中央和侧面阵列的传感器元素的布局创建草稿设计。将设计提交到此探测器的制造商。</li>
		<li>通过制造商的反馈, 为自定义探头创建详细的设计, 包括改进传感器频率特性和几何图形 (图 2)。 注意: 通常, 自定义探头的制造商可以设计电子系统、探头外壳和探头连接器, 以兼容特定类型的超声波成像系统。制造商还可以包括一个操作模式开关 (硬件), 以确定哪一套128元素是由超声成像系统解决。在成像模式中, 对中心阵列进行了处理;在跟踪模式中, 将对侧面数组进行寻址。</li>
	</ol>
</li>
	<li>跟踪针<ol>
<li>选择一种光纤超声水听器, 其包括 Pérot 腔的单模光纤, 其远端 (外径): 150 µm)。 注: 水听器, 包括一个单模光纤与法 Pérot 腔在远端 (OD: 150 µm), 是可利用的商业。近端到远端, 通常用于电信的光纤有一个包覆层 (od: 125 µm), 缓冲层 (od: 250 µm) 和夹克 (od: 900 µm)。</li>
		<li>使用手术刀, 部分删除900微米夹克沿长度的光纤水听器, 接近其远端, 以暴露的缓冲层, 直到水听器可以适合针套管内。 注: 对于机械鲁棒性, 在鲁尔连接器的光纤电缆的截面上保留保护缓冲层/护套是很有用的。注意处理后的纤维易碎的部分后, 夹克被删除, 之前, 它是由针套管保护。</li>
		<li>将医用针水平贴到手动水平平移阶段, 用立体显微镜可视化针尖, 显微镜的光轴水平和垂直对准针。如有必要, 旋转针的轴线, 使针的斜角表面可以看到与显微镜。</li>
		<li>随着针的远端, 显微镜, 插入光纤超声接收机通过套管的杜希-Borst 手枪适配器, 随后通过鲁尔连接器的针, 直到传感区的水听机只是针的斜角表面的近端。在这个阶段, 手枪适配器不应该连接到针。将水听器贴到翻译阶段 (聚酰亚胺胶带工作良好), 以避免其在针内运动。</li>
		<li>用聚酰亚胺胶带将水听器贴到翻译阶段, 以避免设备在针内移动。</li>
		<li>将20微升的吸管垂直贴在垂直平移阶段, 尖端朝下向下, 同时使用水平和垂直平移阶段定位微尖端, 直到它与光纤水听器相邻, 大约0.5 毫米近端到远端的传感区。</li>
		<li>在微的近端放置一滴光学粘合剂, 并调整针, 允许从微尖端到光纤超声接收机的直接路径。</li>
		<li>然后使用10毫升注射器在微的近端施加压力, 逐渐将粘合剂从远端剥离到光纤超声接收机中, 注意避免将胶粘剂应用于传感区或阻断套管, 并用紫外线照射针尖, 直到光学粘合剂固化。</li>
	</ol>
</li>
</ol>2. 系统集成
<ol><li>将水听器连接到其光学控制台。 注: 提供模拟电压信号与所接收压力成正比的光学控制台可在商业上使用。</li>
	<li>将自定义超声波成像探头连接到超声波控制台。</li>
	<li>执行交叉收购 B 型超声图像和编码超声脉冲跟踪10,14。对于 B 型超声图像采集, 对中心阵元进行脉冲回波发射接收序列。使用硬件开关控制是否访问侧面阵列元素或中央数组元素。</li>
	<li>将水听器信号和定时信号数字化, 同时使用数据采集 (数据采集) 卡同时进行超声波传输。</li>
	<li>处理并显示从脉冲回波发送接收序列获取的信号, 获得 B 型超声图像。此外, 处理和显示水听器信号, 以本地化光纤超声接收机相对于自定义探头。对于后一个任务, 这些算法被描述为夏et al。12,14
</li>
	<li>将针尖位置覆盖到 B 型超声图像上。要将3D 跟踪信息显示在2D 超声图像显示上, 可以用十字表示针尖 (横向和深度坐标) 的位置;成像平面的平面距离和侧面, 其大小和颜色分别为这个十字。</li>
</ol>3. 临床前验证
<ol><li>使用超声波成像探头上的开关选择操作模式。</li>
	<li>在自定义超声成像探头中添加超声波凝胶。</li>
	<li>通过添加水模拟羊水, 制备胎儿超声幻影。</li>
	<li>采用 B 型超声成像技术, 将幻影中的羊水识别为插入靶。 注: 插入目标将取决于上下文;它可以包括一个特定区域的组织用于诊断或治疗在临床过程中, 或指定的位置在成像幻影模仿组织区域。</li>
	<li>向插入目标插入针。在插入过程中, 在操作模式 (成像和跟踪) 之间交替使用自定义探测器上的开关。</li>
</ol>

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作者声明没有利益冲突。

在这里, 我们演示了如何使用自定义的超声成像探头和光纤水听器集成在一个针头, 3D 超声波跟踪。从临床翻译的角度来看, 本研究开发的自定义探针的几个方面是有吸引力的。它紧凑的大小是非常适合使用的小空间, 如腋窝, 机动笨重的3D 成像探头是挑战。这里提出的3D 超声波跟踪的一个局限性是, 需要手动切换以在成像和跟踪模式之间交替。在将来的实现中, 这种切换可以直接由超声成像系统进行。
光纤水听器适用于超声针跟踪。它的高度小型化和灵活性, 使其融入到小的横向尺寸的医疗设备。其广泛的频率带宽16允许与不同的临床超声探头兼容。此外, 它的 omnidirectionality16允许跟踪针, 插入在很大范围的角度。最后, 它对干扰从 em 领域和金属对象的免疫力使它更适合于临床设置与 em 跟踪对比。为达到更大的超声检测灵敏度, 可在未来17采用平凹法 Pérot 腔。最终, 超声波跟踪可以与其他方式结合在一个单一的光纤, 如反射光谱18,19,20,21,22,23、拉曼光谱24、光学相干层析成像 25、26、光声成像27、28、29、30,31,32,33。
超声跟踪具有与超声成像共享的局限性。首先, 组织异质性会对超声跟踪产生负面影响;在前一项研究中, 通过数值模拟表明, 组织声音速度的空间变化将会降低跟踪精度.第二, 高度反射到超声波的解剖结构, 如骨结构或空气空洞, 可能与超声追踪不相容。在今后的研究中, 用其他成像方式获得的针尖位置, 如3D 旋转 C 臂计算 X 射线层析成像, 可用于评估体内非均质组织中3D 超声跟踪的准确性。
尽管最近在超声成像方面有所进展, 但在这种模式的指导下, 精确跟踪和有效操纵医疗器械仍然具有挑战性, 即使对于专家从业者也是如此。如图所示, 外部超声探头与医疗器械之间的主动通信可以提高程序的安全性和效率。这些改进可以极大地促进采用超声成像代替 X 射线透视在几个临床情况下, 如脊柱插入的介入疼痛管理。本研究开发的系统使3D 超声跟踪和2D 超声成像与紧凑型超声探头。通过在当前临床工作流程中精确定位针尖, 可以改善超声引导下的微创手术。

在许多超声引导下的微创手术中, 对侵入性医疗器械进行准确有效的定位是非常理想的。这些程序是在临床环境中遇到的, 如区域麻醉和介入疼痛管理1, 介入肿瘤学2和胎儿医学3。医疗器械尖端的可视化对超声成像有挑战性。在平面插入过程中, 当插入角陡峭时, 针头的能见度往往很差。此外, 在平面外的插入, 针轴可以被误解为针尖。当针尖不是超声可见, 它可能会导致并发症的破坏关键组织结构。
许多方法可用于在超声成像过程中定位医疗器械, 但与当前临床工作流程相适应的可靠方案是非常理想的。回声表面可用于提高能见度在陡峭的角度在平面插入4。电磁跟踪系统可以在平面外插入过程中使用, 但电磁场干扰会严重降低其准确度。3D 超声成像可以提高医疗器械在某些心脏和胎儿的程序, 当他们被液体包围5的能见度。然而, 3D 超声成像不广泛用于针引导, 部分原因是由于复杂的图像解释。
超声跟踪是提高医疗器械能见度的一种方法,6、7、8、9、10、11、12 ,13,14。在超声跟踪的情况下, 该医疗器械有一个嵌入式超声传感器或发射机, 主动与外部超声成像探头进行通信。医疗器械的位置可以从测量的超声飞行时间间隔内的嵌入式超声传感器/发射机和不同的传感器元件的探头。迄今为止, 超声跟踪仅限于平面跟踪, 这大大限制了它的临床应用。
这里演示了如何使用自定义的超声成像探头和在针的套管内贴上的光纤水听器来进行3D 超声波跟踪 (图 1)。该自定义探头是由作者设计和外部制造的, 包括一个中央阵列的传感器元件和四侧阵列。中心阵列用于2D 超声成像;侧面阵列, 用于3D 针尖跟踪与光纤超声接收机的配合。分析了光纤超声接收机在针套管内的定位和贴接方式, 如何在台式上测量系统的跟踪精度, 以及如何进行临床验证。

<table><tbody><tr><td>Ultrasound imaging system</td><td>BK ultrasound (ultrasonix)</td><td>SonixMDP</td><td></td></tr><tr><td>Custom ultrasound probe</td><td>Vermon</td><td></td><td></td></tr><tr><td>Spinal needle&amp;nbsp;</td><td>Terumo</td><td></td><td>20 gauge</td></tr><tr><td>Fibre-optic hydrophone</td><td>Precision Acoustics</td><td></td><td></td></tr><tr><td>Fibre-optic stripping tool&amp;nbsp;</td><td>Thorlabs</td><td>FTS4</td><td></td></tr><tr><td>Stereo microscope&amp;nbsp;</td><td>Leica Microsystems&amp;nbsp;</td><td>Z16APO</td><td></td></tr><tr><td>Tuohy-Borst Sidearm adapter&amp;nbsp;</td><td>Cook Medical</td><td>PTBYC-RA</td><td></td></tr><tr><td>Pipette&amp;nbsp;&amp;nbsp;</td><td>Eppendorf</td><td></td><td>100 mL</td></tr><tr><td>Micropipette tip&amp;nbsp;</td><td>Eppendorf</td><td></td><td>20 &amp;micro;L</td></tr><tr><td>Ultraviolet optical adhesive&amp;nbsp;</td><td>Norland Products</td><td>NOA81</td><td></td></tr><tr><td>Syringe</td><td>Terumo</td><td></td><td>&amp;nbsp;10 mL</td></tr><tr><td>Ultraviolet light source&amp;nbsp;</td><td>Norland Products</td><td>Opticure 4 Light Gun</td><td></td></tr><tr><td>Data acquisiton card&amp;nbsp;</td><td>National Instruments</td><td>USB-5132</td><td></td></tr><tr><td>Articulated arm&amp;nbsp;</td><td>CIVCO</td><td>811-002</td><td></td></tr><tr><td>Thiopental sodium&amp;nbsp;</td><td>Novartis Animal Health UK&amp;nbsp;</td><td></td><td>Thiovet</td></tr><tr><td>Isoflurane</td><td>Merial Animal Health</td><td></td><td>Isoflurane-Vet</td></tr><tr><td>Ocular lubricant</td><td>Allergan, Marlow, UK</td><td></td><td>Lacri-Lube</td></tr><tr><td>Skin lubricant</td><td>Adams Healthcare, Garforth, UK</td><td></td><td>Hibitane 2%</td></tr></tbody></table>

three-dimensional ultrasonic needle tip tracking with a fiber-optic ultrasound receiver

超声通常用于指导微创手术, 但可视化的医疗器械往往挑战这种成像方式。当可视化丢失时, 医疗器械会对关键组织结构造成创伤。本文给出了一种在超声图像导引过程中跟踪针尖的方法。这种方法涉及使用一种光纤超声接收器, 它被贴在医用针的套管内, 与外超声探头进行超声交流。该自定义探头包括中央传感器元件阵列和侧面元件阵列。除了由中央阵列提供的常规二维 (2D) B 型超声成像外, 侧面阵列还提供了三维 (3D) 针尖跟踪。对于 B 型超声成像, 采用电子波束形成的标准发射接收序列。对于超声波跟踪, 通过水听器传感器接收4侧阵列的 Golay 编码超声传输, 随后对接收到的信号进行解码, 以识别针尖在超声成像方面的空间位置。探针。作为对该方法的初步验证, 对针/水听器对的插入在临床现实环境中进行。这种新的超声成像/跟踪方法与目前的临床工作流程相适应, 在平面和平面外针插入过程中提供了可靠的设备跟踪。

动物实验是根据英国家庭办公室章程和指南为动物操作 (科学规程) 行动 (1986)。根据英国内政部有关动物福利的准则, 将绵羊安置在该处;这些实验是在70/7408 的家庭办公室项目许可下进行的, 题为 "干细胞和基因转移的产前治疗"。英国伦敦大学学院和皇家兽医学院动物福利伦理审查委员会提供了绵羊实验的伦理学批准。
随着伦理学的批准, 一只怀孕的绵羊用于临床前的体内验证。在接受阴道孕酮栓2星期后, 母羊是时间交配诱发排卵, 如大卫等 al所描述的。34在130天的妊娠期, 一只怀孕的母羊一夜之间饿死, 一只怀孕的同伴母羊。然后, 母羊进行全身麻醉, 经硫喷妥钠20毫克-1的静脉注射, 并维持 2-2. 5% 异氟醚在氧气插管后通过呼吸机。正确的气管插管是通过双侧听肺部证实的。通过对角膜反射的评估来证实麻醉。用舌或耳上的饱和监测仪连续测量氧饱和度。母羊被放在她的背部在半 recundancy 和鼻胃管被通过, 以缓解胃内容的通过。眼部润滑剂被应用于眼睛, 以保持其湿润。羊毛修剪后, 母羊的腹部用皮肤消毒剂双擦洗。采用无菌偶联凝胶对腹部进行超声检查, 确定母羊34胎龄, 评估胎儿假谎。手术结束时, 使用过量的硫喷妥钠 (静脉注射40毫克-1 ), 人道地杀死了该动物。
医生 (A.L.D.) 确认脐带为目标。在子宫腔内插入一根针, 并沿轨迹进行跟踪, 达到15毫米的平面距, 深度为38毫米 (图 3)。Golay 编码提高了信噪比, 与传统的双极励磁相比增加了7.5 倍 (图 3B)。3D 履带针尖位置被覆盖在2D 超声图像上使用交叉宽度指示出平面距离和颜色指示成像 (步骤 2.6) (图 3C)。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/57207/57207fig1.jpg"> 图 1: 系统概述.超声 (美国) 成像/跟踪探头既允许2D 美国成像和3D 针跟踪。它由美国扫描仪驱动, 它提供对跟踪元件传输的控制。开关允许换能器元件的电子选择在两种操作模式之间交替: 成像与中央阵列和跟踪与侧面阵列。光纤水听器 (FOH) 超声波接收机, 定位在20G 针的腔内, 接收来自侧面阵列的传输。T/R: 发送/接收;直线触发;FT: 帧触发器;个人计算机: 个人计算机;数据采集卡。这个数字和标题是转载的, 由夏, W等人的许可。14.<a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/57207/57207fig1large.jpg" target="_blank">请点击这里查看这个数字的更大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/57207/57207fig2.jpg"> 图 2: 自定义超声成像探头的传感器元件布局.具有128个元素和一个声学透镜的中央阵列使我们能够成像。侧阵列, 每行32个元素, 总共有128个元素, 可实现3D 针跟踪。这个数字和标题是转载的, 由夏, W等人的许可。14.<a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/57207/57207fig2large.jpg" target="_blank">请点击这里查看这个数字的更大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/57207/57207fig3.jpg"> 图 3:在体内进行3D 跟踪的针插入。(A) 在怀孕绵羊的子宫腔内插入时所获得的跟踪针尖位置 (圆圈: P1-P6)。(B) 跟踪信号的信噪比 (信噪比) (成像平面: X = 0)。(C) 将3的履带位置覆盖在与中央阵列一起获得的2D 美国图像上。每个十字的端到端长度对应于平面外距离;颜色 (红色/黄色) 对应于成像平面的侧面。主要解剖特征用轮廓 (右) 描述。S: 皮肤;PF: 经皮脂肪;子宫壁;羊膜: 羊水;脐带;胎儿腹部。这个数字和标题是转载的, 由夏, W等人的许可。14.<a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/57207/57207fig3large.jpg" target="_blank">请点击这里查看这个数字的更大版本.</a>

Watch this Scientific Journal Video about Three-Dimensional Ultrasonic Needle Tip Tracking with a Fiber-Optic Ultrasound Receiver at JoVE.com

Three-Dimensional Ultrasonic Needle Tip Tracking with a Fiber-Optic Ultrasound Receiver

基于光纤超声接收机的三维超声针尖跟踪

在许多超声引导下的微创手术中, 准确有效地显示侵入性医疗器械是极其重要的。本文提出了一种相对于超声成像探针定位针尖空间位置的方法。

Ultrasound is frequently used for guiding minimally invasive procedures, but visualizing medical devices is often challenging with this imaging modality. ...

three-dimensional-ultrasonic-needle-tip-tracking-with-fiber-optic

Universidad de Cartagena UDEC

Research

JoVE Journal

Engineering

10.3K 次观看.  University College London. 在许多超声引导下的微创手术中, 准确有效地显示侵入性医疗器械是极其重要的。本文提出了一种相对于超声成像探针定位针尖空间位置的方法。

视频: Three-Dimensional Ultrasonic Needle Tip Tracking with a Fiber-Optic Ultrasound Receiver

Three-dimensional Optical-resolution Photoacoustic Microscopy

Optical microscopy, providing valuable insights at the cellular and organelle levels, has been widely recognized as an enabling biomedical technology. As the mainstays of in vivo three-dimensional (3-D) optical microscopy, single-/multi-photon fluorescence microscopy and optical coherence tomography (OCT) have demonstrated their extraordinary sensitivities to fluorescence and optical scattering contrasts, respectively. However, the optical absorption contrast of biological tissues, which encodes essential physiological/pathological information, has not yet been assessable. 
The emergence of biomedical photoacoustics has led to a new branch of optical microscopy optical-resolution photoacoustic microscopy (OR-PAM)1, where the optical irradiation is focused to the diffraction limit to achieve cellular1 or even subcellular2 level lateral resolution. As a valuable complement to existing optical microscopy technologies, OR-PAM brings in at least two novelties. First and most importantly, OR-PAM detects optical absorption contrasts with extraordinary sensitivity (i.e., 100%). Combining OR-PAM with fluorescence microscopy3 or with optical-scattering-based OCT4 (or with both) provides comprehensive optical properties of biological tissues. Second, OR-PAM encodes optical absorption into acoustic waves, in contrast to the pure optical processes in fluorescence microscopy and OCT, and provides background-free detection. The acoustic detection in OR-PAM mitigates the impacts of optical scattering on signal degradation and naturally eliminates possible interferences (i.e., crosstalks) between excitation and detection, which is a common problem in fluorescence microscopy due to the overlap between the excitation and fluorescence spectra. 
Unique for optical absorption imaging, OR-PAM has demonstrated broad biomedical applications since its invention, including, but not limited to, neurology5, 6, ophthalmology7, 8, vascular biology9, and dermatology10. In this video, we teach the system configuration and alignment of OR-PAM as well as the experimental procedures for in vivo functional microvascular imaging.

Optical-resolution photoacoustic microscopy (OR-PAM) is an emerging technology capable of imaging optical absorption contrasts in vivo with cellular resolution and sensitivity. Here, we provide a visualized instruction on the experimental protocols of OR-PAM, including system configuration, system alignment, typical in vivo experimental procedures, and functional imaging schemes.

三维光学分辨率的光声显微镜

Optical microscopy, providing valuable insights at the cellular and organelle levels, has been widely recognized as an enabling biomedical technology. As ...

科研

生物工程

Wideband Optical Detector of Ultrasound for Medical Imaging Applications

Optical sensors of ultrasound are a promising alternative to piezoelectric techniques, as has been recently demonstrated in the field of optoacoustic imaging. In medical applications, one of the major limitations of optical sensing technology is its susceptibility to environmental conditions, e.g. changes in pressure and temperature, which may saturate the detection. Additionally, the clinical environment often imposes stringent limits on the size and robustness of the sensor. In this work, the combination of pulse interferometry and fiber-based optical sensing is demonstrated for ultrasound detection. Pulse interferometry enables robust performance of the readout system in the presence of rapid variations in the environmental conditions, whereas the use of all-fiber technology leads to a mechanically flexible sensing element compatible with highly demanding medical applications such as intravascular imaging. In order to achieve a short sensor length, a pi-phase-shifted fiber Bragg grating is used, which acts as a resonator trapping light over an effective length of 350 &#181;m. To enable high bandwidth, the sensor is used for sideway detection of ultrasound, which is highly beneficial in circumferential imaging geometries such as intravascular imaging. An optoacoustic imaging setup is used to determine the response of the sensor for acoustic point sources at different positions.

Optical detection of ultrasound is impractical in many imaging scenarios because it often requires stable environmental conditions. We demonstrate an optical technique for ultrasound sensing in volatile environments with miniaturization and sensitivity levels appropriate for optoacoustic imaging in restrictive scenarios, e.g. intravascular applications.

宽带超声医疗成像应用，光学检测仪

Optical sensors of ultrasound are a promising alternative to piezoelectric techniques, as has been recently demonstrated in the field of optoacoustic ...

3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles

The objective of this video protocol is to discuss how to perform and analyze a three-dimensional fluorescent orbital particle tracking experiment using a modified two-photon microscope1. As opposed to conventional approaches (raster scan or wide field based on a stack of frames), the 3D orbital tracking allows to localize and follow with a high spatial (10 nm accuracy) and temporal resolution (50 Hz frequency response) the 3D displacement of a moving fluorescent particle on length-scales of hundreds of microns2. The method is based on a feedback algorithm that controls the hardware of a two-photon laser scanning microscope in order to perform a circular orbit around the object to be tracked: the feedback mechanism will maintain the fluorescent object in the center by controlling the displacement of the scanning beam3-5. To demonstrate the advantages of this technique, we followed a fast moving organelle, the lysosome, within a living cell6,7. Cells were plated according to standard protocols, and stained using a commercially lysosome dye. We discuss briefly the hardware configuration and in more detail the control software, to perform a 3D orbital tracking experiment inside living cells. We discuss in detail the parameters required in order to control the scanning microscope and enable the motion of the beam in a closed orbit around the particle. We conclude by demonstrating how this method can be effectively used to track the fast motion of a labeled lysosome along microtubules in 3D within a live cell. Lysosomes can move with speeds in the range of 0.4-0.5 &#181;m/sec, typically displaying a directed motion along the microtubule network8.

In this video protocol we track - at high speed and in three dimensions - fluorescently labeled lysosomes within living cells, using the orbital tracking method in a modified two-photon microscope.

3d轨道跟踪的改进双光子显微镜之应用:细胞内的囊泡的跟踪

The objective of this video protocol is to discuss how to perform and analyze a three-dimensional fluorescent orbital particle tracking experiment using a ...

Universal Hand-held Three-dimensional Optoacoustic Imaging Probe for Deep Tissue Human Angiography and Functional Preclinical Studies in Real Time

The exclusive combination of high optical contrast and excellent spatial resolution makes optoacoustics (photoacoustics) ideal for simultaneously attaining anatomical, functional and molecular contrast in deep optically opaque tissues. While enormous potential has been recently demonstrated in the application of optoacoustics for small animal research, vast efforts have also been undertaken in translating this imaging technology into clinical practice. We present here a newly developed optoacoustic tomography approach capable of delivering high resolution and spectrally enriched volumetric images of tissue morphology and function in real time. A detailed description of the experimental protocol for operating with the imaging system in both hand-held and stationary modes is provided and showcased for different potential scenarios involving functional and molecular studies in murine models and humans. The possibility for real time visualization in three dimensions along with the versatile handheld design of the imaging probe make the newly developed approach unique among the pantheon of imaging modalities used in today&#8217;s preclinical research and clinical practice.

We provide herein a detailed description of the experimental protocol for imaging with a newly developed hand-held optoacoustic (photoacoustic) system for three-dimensional functional and molecular imaging in real time. The demonstrated powerful performance and versatility may define new application areas of the optoacoustic technology in preclinical research and clinical practice.

通用型手持式三维光声成像探测深部组织人力造影和功能的临床前研究中实时

The exclusive combination of high optical contrast and excellent spatial resolution makes optoacoustics (photoacoustics) ideal for simultaneously ...

Multiplexing Focused Ultrasound Stimulation with Fluorescence Microscopy

By focusing low-intensity ultrasound pulses that penetrate soft tissues, LIPUS represents a promising biomedical technology to remotely and safely manipulate neural firing, hormonal secretion and genetically-reprogrammed cells. However, the translation of this technology for medical applications is currently hampered by a lack of biophysical mechanisms by which targeted tissues sense and respond to LIPUS. A suitable approach to identify these mechanisms would be to use optical biosensors in combination with LIPUS to determine underlying signaling pathways. However, implementing LIPUS to a fluorescence microscope may introduce undesired mechanical artefacts due to the presence of physical interfaces that reflect, absorb and refract acoustic waves. This article presents a step-by-step procedure to incorporate LIPUS to commercially-available upright epi-fluorescence microscopes while minimizing the influence of physical interfaces along the acoustic path. A simple procedure is described to operate a single-element ultrasound transducer and to bring the focal zone of the transducer into the objective focal point. The use of LIPUS is illustrated with an example of LIPUS-induced calcium transients in cultured human glioblastoma cells measured using calcium imaging.

Low-Intensity Pulsed Ultrasound Stimulation (LIPUS) is a modality for non-invasive mechanical stimulation of endogenous or engineered cells with high spatial and temporal resolution. This article describes how to implement LIPUS to an epi-fluorescence microscope and how to minimize acoustic impedance mismatch along the ultrasound path to prevent unwanted mechanical artefacts.

荧光显微镜对聚焦超声的刺激

By focusing low-intensity ultrasound pulses that penetrate soft tissues, LIPUS represents a promising biomedical technology to remotely and safely ...

工程学

Evaluating Targeting Accuracy in the Focal Plane for an Ultrasound-guided High-intensity Focused Ultrasound Phased-array System

Phased arrays are increasingly used as high-intensity focused ultrasound (HIFU) transducers in the existing extracorporeal ultrasound-guided HIFU (USgHIFU) systems. The HIFU transducers in such systems are usually spherical in shape with a central hole where a US imaging probe is mounted and can be rotated. The image on the plane of treatment can be reconstructed through the image sequence acquired during the rotation of the probe. Therefore, the treatment plan can be made on the reconstructed images. In order to evaluate the targeting accuracy in the focal plane of such systems, the protocol of a method using a bovine muscle and marker-embedded phantom is described. In the phantom, four solid balls at the corners of a square resin model serve as the reference markers in the reconstructed image. The target should be moved so that both its center and the center of the square model can coincide according to their relative positions in the reconstructed image. Swine muscle with a thickness of about 30 mm is placed above the phantom to mimic the beam path in clinical settings. After sonication, the treatment plane in the phantom is scanned and the boundary of the associated lesion is extracted from the scanned image. The targeting accuracy can be evaluated by measuring the distance between the centers of target and lesion, as well as three derivative parameters. This method cannot only evaluate the targeting accuracy of the target consisting of multiple focal spots rather than a single focal spot in a clinically relevant beam path of the USgHIFU phased-array system, but it can be also used in the preclinical evaluation or regular maintenance of USgHIFU systems configured with phased-array or self-focused HIFU transducer.

This study describes a protocol to evaluate the targeting accuracy in the focal plane of an ultrasound-guided high-intensity focused ultrasound phased-array system.

超声引导高强度聚焦超声相控阵系统焦平面定位精度的评价

Phased arrays are increasingly used as high-intensity focused ultrasound (HIFU) transducers in the existing extracorporeal ultrasound-guided HIFU ...

An Experimental Protocol for Assessing the Performance of New Ultrasound Probes Based on CMUT Technology in Application to Brain Imaging

The possibility to perform an early and repeatable assessment of imaging performance is fundamental in the design and development process of new ultrasound (US) probes. Particularly, a more realistic analysis with application-specific imaging targets can be extremely valuable to assess the expected performance of US probes in their potential clinical field of application.
The experimental protocol presented in this work was purposely designed to provide an application-specific assessment procedure for newly-developed US probe prototypes based on Capacitive Micromachined Ultrasonic Transducer (CMUT) technology in relation to brain imaging.
The protocol combines the use of a bovine brain fixed in formalin as the imaging target, which ensures both realism and repeatability of the described procedures, and of neuronavigation techniques borrowed from neurosurgery. The US probe is in fact connected to a motion tracking system which acquires position data and enables the superposition of US images to reference Magnetic Resonance (MR) images of the brain. This provides a means for human experts to perform a visual qualitative assessment of the US probe imaging performance and to compare acquisitions made with different probes. Moreover, the protocol relies on the use of a complete and open research and development system for US image acquisition, i.e. the Ultrasound Advanced Open Platform (ULA-OP) scanner.
The manuscript describes in detail the instruments and procedures involved in the protocol, in particular for&#160;the calibration, image acquisition and registration of US and MR images. The obtained results prove the effectiveness of the overall protocol presented, which is entirely open (within the limits of the instrumentation involved), repeatable, and covers the entire set of acquisition and processing activities for US images.

The development of new ultrasound (US) probes based on Capacitive Micromachined Ultrasonic Transducer (CMUT) technology requires an early realistic assessment of imaging capabilities. We describe a repeatable experimental protocol for US image acquisition and comparison with magnetic resonance images, using an ex vivo bovine brain as an imaging target.

基于 CMUT 技术的新型超声探头在脑成像中的应用研究

The possibility to perform an early and repeatable assessment of imaging performance is fundamental in the design and development process of new ...

Hand-held Clinical Photoacoustic Imaging System for Real-time Non-invasive Small Animal Imaging

Translation of photoacoustic imaging into the clinic is a major challenge. Handheld real-time clinical photoacoustic imaging systems are very rare. Here, we report a combined photoacoustic and clinical ultrasound imaging system by integrating an ultrasound probe with light delivery for small animal imaging. We demonstrate this by showing sentinel lymph node imaging in small animals along with minimally invasive real-time needle guidance. A clinical ultrasound platform with access to raw channel data allows the integration of photoacoustic imaging leading to a handheld real-time clinical photoacoustic imaging system. Methylene blue was used for sentinel lymph node imaging at 675 nm wavelength. Additionally, needle guidance with dual modal ultrasound and photoacoustic imaging was shown using the imaging system. Depth imaging of up to 1.5 cm was demonstrated with a 10 Hz laser at a photoacoustic imaging frame rate of 5 frames per second.

A clinical handheld photoacoustic imaging system will be demonstrated for real-time non-invasive small animal imaging.

手持式实时非侵入性小动物成像的临床光声成像系统

Translation of photoacoustic imaging into the clinic is a major challenge. Handheld real-time clinical photoacoustic imaging systems are very rare. Here, ...

A Protocol for Real-time 3D Single Particle Tracking

Real-time three-dimensional single particle tracking (RT-3D-SPT) has the potential to shed light on fast, 3D processes in cellular systems. Although various RT-3D-SPT methods have been put forward in recent years, tracking high speed 3D diffusing particles at low photon count rates remains a challenge. Moreover, RT-3D-SPT setups are generally complex and difficult to implement, limiting their widespread application to biological problems. This protocol presents a RT-3D-SPT system named 3D Dynamic Photon Localization Tracking (3D-DyPLoT), which can track particles with high diffusive speed (up to 20 &#181;m2/s) at low photon count rates (down to 10 kHz). 3D-DyPLoT employs a 2D electro-optic deflector (2D-EOD) and a tunable acoustic gradient (TAG) lens to drive a single focused laser spot dynamically in 3D. Combined with an optimized position estimation algorithm, 3D-DyPLoT can lock onto single particles with high tracking speed and high localization precision. Owing to the single excitation and single detection path layout, 3D-DyPLoT is robust and easy to set up. This protocol discusses how to build 3D-DyPLoT step by step. First, the optical layout is described. Next, the system is calibrated and optimized by raster scanning a 190 nm fluorescent bead with the piezoelectric nanopositioner. Finally, to demonstrate real-time 3D tracking ability, 110 nm fluorescent beads are tracked in water.

This protocol details the construction and operation of a real-time 3D single particle tracking microscope capable of tracking nanoscale fluorescent probes at high diffusive speeds and low photon count rates.

一种实时3D 单粒子跟踪协议

Real-time three-dimensional single particle tracking (RT-3D-SPT) has the potential to shed light on fast, 3D processes in cellular systems. Although ...

Three-Dimensional Finger Motion Tracking during Needling: A Solution for the Kinematic Analysis of Acupuncture Manipulation

Three-dimensional (3D) motion tracking has been used in many fields, such as the researches of sport and medical skills. This experiment aimed to use 3D motion tracking technology to measure the kinematic parameters of the joints of fingers during acupuncture manipulation (AM) and establish three technical indicators "amplitude, velocity and time". This method can reflect the operation characteristics of AM and provide quantitative parameters along three axes of multiple finger joints. The current evidence shows that the method has great potential for future applications such as the study of the dose-effect relationship of acupuncture, teaching, and learning of AM, and the measurement and preservation of famous acupuncturists' AM.

This experimental method describes a solution for the kinematic analysis of acupuncture manipulation with three-dimensional finger motion tracking technology.

针刺过程中的三维手指运动跟踪:针灸操作运动学分析的解决方案

Three-dimensional (3D) motion tracking has been used in many fields, such as the researches of sport and medical skills. This experiment aimed to use 3D ...