这项工作得到了国家政府和欧洲联盟的部分支持, 通过 ENIAC JU 项目 DeNeCoR 324257 号赠款协议。作者要感谢 Prof. 乔瓦尼 Magenes, Prof. 皮托尔托利和 Dr. Giosuè Caliano 为他们宝贵的支持, 监督和有见地的意见, 使这项工作成为可能。我们也感谢 Prof. 圣埃吉迪奥和他的小组 (BCC 实验室), 连同基金会依 Neurologico c Mondino, 提供了运动跟踪和 mr 仪器, 并为 Germani 先生收购。最后, 我们要感谢 Dr. 尼可莱塔 Caramia, Dr. Dallai 和 Ms. 芭芭拉 Mauti 为他们的宝贵技术支持和 Mr. 沃尔特沃尔皮提供牛脑。

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作者宣布, 他们没有竞争的金融利益。

在文献中介绍了一些与所提出的协议相似或相关的技术。这些技术也基于实际目标的使用, 包括固定的动物或尸体的大脑, 但它们主要是为测试各种类型的数字注册方法。
然而, 此处描述的协议具有特定的目的, 即在开发的早期阶段对美国探测器进行不同的配置测试, 因此, 它满足了收购的可再现性的基本要求, 即在同一生物标本和具有可比的姿势。该协议介绍了上述现有技术的许多方面, 并以不同的方式将它们组装在一起。
在协议设计和实验中所吸取的教训中, 校准程序是目前最关键的方面。尽管被采取的许多改善, 整个转换集合的整体空间错误在定标以后是当前在命令 1-1. 5 毫米。这样的错误不是由于运动跟踪器的精确度 (它的记录精度是 0.3 mm), 而是由于在生物标本上获取精确的空间读数的难度, 这就保留了一定的灵活性。
另一方面, 在我们的经验中, 同步的时间精度并不是一个关键的方面。事实上, 运动跟踪器的位置数据的获取率是一个比人的手的动作更大的数量级, 试图达到一个稳定的姿态。由于这个原因, 在协议中计算的时间平均值是为了获得额外的精确性。另一个特别有效的方面是虚拟姿势的定义。在进行的实验中, 多亏了 real-time 的视觉跟踪程序, 操作员可以在不费力和支持机械结构的情况下, 为三美国探针中的每一个十二虚拟体式的图像进行采集。
对该协议的一个可能的修改, 将在未来采用, 是使用不同的和改进的校准方法, 这应该是基于一个更紧密的循环和空间转换的反馈。在目前的形式, 实际上, 协议需要大量的空间读数后处理, 以计算转换矩阵。虽然此活动可以在数十分钟内执行, 并且不需要将实验置于离线状态, 但此后处理提供了在执行校准时无法立即可视化的结果。在这方面, 一个增强的和可能 real-time 的视觉反馈的校准得到了很大的帮助, 以实现更高的精确度。
对于议定书的实际执行而言, 必须有合理开放的工具, 并允许多种必要的集成。例如, 在这种情况下, 通过 ULA-OP 系统提供的内部计时数据的访问来确保来自不同来源的信号的同步的实际可能性, 对于校准和图像后处理活动都是至关重要的。
另一个重要因素是软件。虽然实验不需要主要的软件工具, 但许多 c++ 和 MATLAB 例程, 加上 Python-based 模块的 Paraview, 证明了重要的一些关键任务, 如校准, 运动跟踪反馈的预定义3D 图像重建的构成和后处理。再次, 获得这些仪器所产生的低级数据对于创建这些软件组件非常重要。
最后, 选择合适的成像目标是非常重要的。考虑到合成幻影的实现的几个替代方案事先被认为, 根据我们的经验, 所有这些选择似乎是不理想的, 相比之下, 在福尔马林固定的牛脑 cost-effective 选择。这一目标确保了更好的现实主义, 并以适当的照顾, 无限期的保存随着时间的推移。
最后, 以3D 多模态图像数据集作为一个永久性的相关结果, 得出的实验结果, 在我们看来, 是一个有效的技术集成策略的产物, 需要分段组装,通过仔细分析隐含的许多方面, 并就所涉及的程序和文书进行设计。

<table><tbody><tr><td>ULA-OP</td><td>University of Florence</td><td>N/A</td><td>Ultrasound imaging research system</td></tr><tr><td>3D imaging piezeoelectric probe</td><td>Esaote s.p.a.</td><td>9600195000</td><td>Mechanically-swept 3D ultrasound probe, model BL-433</td></tr><tr><td>Linear-array piezoelectric probe</td><td>Esaote s.p.a.</td><td>122001100</td><td>Ultrasound linear array probe, model LA-533</td></tr><tr><td>CMUT probe</td><td>University Roma Tre</td><td>N/A</td><td>Ultrasound linear array probe based on CMUT technology</td></tr><tr><td>MAGNETOM Skyra 3T MR scanner</td><td>Siemens Healthcare</td><td>N/A</td><td>MR scanner</td></tr><tr><td>Head coil</td><td>Siemens Healthcare</td><td>N/A</td><td>32-channel head coil for MR imaging</td></tr><tr><td>NDI Polaris Vicra</td><td>NDI Medical</td><td>8700335001</td><td>Optical motion tracking system</td></tr><tr><td>Pointer tool</td><td>NDI Medical</td><td>8700340</td><td>Passive pointer tool with 4 reflecting markers</td></tr><tr><td>Clamp-equipped tool</td><td>NDI Medical</td><td>8700399</td><td>Rigid body with 4 reflecting markers and a clamp to be connected to the US probe handle</td></tr><tr><td>Bovine brain</td><td>N/A</td><td>N/A</td><td>Brain of an adult bovine, from food suppliers</td></tr><tr><td>Formalin solution</td><td>N/A</td><td>N/A</td><td>10% buffered formalin solution for bovine brain fixation - CAUTION, formalin is a toxic chemical substance and must be handled with care; specific regulations may also apply (see for instance US OSHA Standard 1910.1048 App A)</td></tr><tr><td>Plastic container for anatomical parts</td><td>N/A</td><td>N/A</td><td>Cilindrical plastic container with lid</td></tr><tr><td>Glass spheres</td><td>N/A</td><td>N/A</td><td>3 mm diameter spheres of Flint glass</td></tr><tr><td>Agar</td><td>N/A</td><td>N/A</td><td>30 g, for phantom preparation</td></tr><tr><td>Glycerine</td><td>AEFFE Farmaceutici</td><td>A908005248</td><td>100 g, for phantom preparation</td></tr><tr><td>Distilled water</td><td>Solbat Gaysol</td><td>8027391000015</td><td>870 g, for phantom preparation</td></tr><tr><td>Beaker</td><td>N/A</td><td>N/A</td><td>Beaker used for the diluition of glycerine and agar in distilled water</td></tr><tr><td>Lysoform</td><td>Lever</td><td>8000680500014</td><td>A benzalkonium chloride and water solution was used for the agar phantom preservation</td></tr><tr><td>Polystyrene mannequin head</td><td>N/A</td><td>N/A</td><td>Polyestirene model which was cutted and used to design the configuration of spheres&#039;patterns</td></tr><tr><td>Green tissue marking dye for histology</td><td>N/A</td><td>N/A</td><td>Colour used to mark the glass spheres&#039; positions on the bovine brain surface</td></tr><tr><td>Yellow enamel</td><td>N/A</td><td>N/A</td><td>Enamel used to colour the glass spheres implanted in the agar phantom</td></tr><tr><td>Water tank</td><td>N/A</td><td>N/A</td><td>50x50x30 cm plastic tank filled with degassed water up to a 15 cm height&amp;nbsp;</td></tr><tr><td>Mechanical arm</td><td>Esaote s.p.a.</td><td>N/A</td><td>Mechanical arm clamped to the water tank border and used to held the probe in fixed positions</td></tr><tr><td>Plate of synthetic resin</td><td>N/A</td><td>N/A</td><td>Plate used as a support for the bovine brain positioning in the water tank</td></tr><tr><td>Sewing threads</td><td>N/A</td><td>N/A</td><td>Sewing thread segments used to immobilize the brain on the resin plate</td></tr><tr><td>Adhesive tape</td><td>N/A</td><td>N/A</td><td>Adhesive tape used to fix the sewing thread extremities onto the resin plate</td></tr><tr><td>Plastic food container</td><td>N/A</td><td>N/A</td><td>Sealed food container used for the agar phantom</td></tr><tr><td>Notebook</td><td>Lenovo</td><td>Z50-70</td><td>Lenovo&amp;nbsp; Z50-70, Intel(R) Core i7-4510U @ 2.0 GHz, 8 GB RAM</td></tr><tr><td>Workstation</td><td>Dell Inc.</td><td>T5810</td><td>Intel(R) Xeon(R) CPU E3-1240v3 @ 3.40 GHz, 16 GB RAM</td></tr><tr><td>Matlab</td><td>The MathWorks</td><td>R2013a</td><td>Software tool, used for space transformation computation and 3D reconstruction from image planes</td></tr><tr><td>Paraview</td><td>Kitware Inc.</td><td>v. 4.4.1</td><td>Open-source software for 3D image processing and visualization</td></tr><tr><td>NDI Toolbox - ToolTracker Utility</td><td>NDI Medical</td><td>v. 4.007.007</td><td>Software for marker position visualization and tracking in the NDI Polaris Vicra measurement volume</td></tr><tr><td>C++ data-logging software</td><td>NDI Medical</td><td>v. 4.007.007</td><td>Software for marker position recording on a text log file</td></tr><tr><td>ULA-OP software&amp;nbsp;</td><td>University of Florence</td><td>N/A</td><td>Software for real-time display and control of the ULA-OP system</td></tr></tbody></table>

an experimental protocol for assessing the performance of new ultrasound probes based on cmut technology in application to brain imaging

对成像性能进行早期和可重复性评估的可能性是新超声 (美国) 探头设计和开发过程中的基础。特别是, 一个更现实的分析与应用特定的成像目标可能是非常宝贵的评估美国探针的预期性能在其潜在的临床应用领域。
在这项工作中提出的实验协议是特意设计的, 以提供一个应用特定的评估程序的新开发的美国探针原型基于电容式微机械超声换能器 (CMUT) 技术的相关脑部成像
该协议结合了使用的牛脑固定在福尔马林作为成像靶, 这既确保了现实主义和重复性的描述的程序, 以及神经导航技术借用从外科。美国探针实际上是连接到一个运动跟踪系统, 获取位置数据, 并使美国图像的叠加, 以参考磁共振 (MR) 的大脑图像。这为人类专家提供了一种方法, 对美国的探针成像性能进行视觉定性评估, 并比较不同探头所做的收购。此外, 该协议依赖于使用一个完整的和开放的研究和开发系统的美国图像采集,即超声高级开放平台 (ULA OP) 扫描仪。
手稿详细描述了协议中涉及的仪器和程序, 特别是对美国和 MR 图像的校准、图像采集和注册。得到的结果证明了整个协议的有效性, 这是完全开放的 (在所涉及的仪器的限制范围内), 可重复, 并涵盖了整个集的收购和处理活动的美国图像。

通过所述协议获得的主要结果是对基于 CMUT 技术的美国探针原型的2D 和3D 成像能力的一种有效且可重复的评估程序的实验验证, 在前瞻性应用于大脑成像.在实现所有描述的协议步骤之后, 专家可以应用可视化软件功能 (例如自由方向切片、子集提取、卷内插、等) 来比较注册的可视化内容美国图像与目标 MR 图像。特别是, 所获得的图像质量, 并与 MRI 的金标准直接比较, 是 CMUT 技术在这一领域的潜力的第一个重要证据。
作为一个可能的可视比较的示例,图 18演示了两个由 CMUT US 探针和压电线性阵列探头分别在叠加到相同的相应切片的体积图像。T2-weighted 先生的形象。T2-weighted 先生的图像证明是最有效的, 从可见性的期望特征, 在这些实验中, 因此被选择为叠加的参考。图中的两个美国图像是以9兆赫的相同频率获得的。如图 18所示, 用 CMUT 探针获得的图像具有更好的分辨率和对比度;此外, 更清晰地定义了突出的可视化功能,沟和回的结构更清楚可见, 这表明 CMUT 探测器的更高灵敏度和更宽的带宽可以提高性能。
<img alt="Figure 18" class="xfigimg" src="/files/ftp_upload/55798/55798fig18.jpg"> 
图 18: 注册美国和 MR 图像切片的叠加.该图显示了通过 CMUT (a、c、e、g) 和压电 (b、d、f、h) 线性阵列探头获得的 mr 和 US 图像的注册。在 (a) 和 (b) 中, 在 MR 空间中显示重建的3D 数据集轮廓, 并突出显示选定的2D 切片。面板 (c、e、g) 和 (d、f、h) 呈现叠加的美国和 MR 切片增加透明度, 以显示两个图像的特征对应。<a href="//ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55798/55798fig18large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
与3D 容量图像相关的进一步的比较示例在图 19和图 20中表示。图 19显示了两个3D 图像, 一个是在体积重建后用压电线性探头获得的, 另一个是用机械扫描压电探头进行3D 成像。图 20显示了用 CMUT 探针获取的图像的3D 体积重构。在所有三种情况下, 大脑皮层的回和沟的3D 结构是清晰可见的, 尽管在与 CMUT 探针获得的体积中, 外部表面更清晰可见, 并且定义得更好。
<img alt="Figure 19" class="xfigimg" src="/files/ftp_upload/55798/55798fig19.jpg"> 
图 19: 用压电探针采集的3D 美国图像.用机械扫描探针 (a、c) 或从平面2D 图像中获得的3D 图像与使用运动跟踪定位数据 (b、d) 的线性探头徒手获得的图象进行比较。在 (a、b) 中, 这些卷的位置显示在 3D MR 图像框中, 使用轮廓。<a href="//ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55798/55798fig19large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 20" class="xfigimg" src="/files/ftp_upload/55798/55798fig20.jpg"> 
图 20: 通过 CMUT 探测器获得的3D 美国重建图像.用 CMUT 探针通过扫描牛脑获得的2D 图像平面被用来重建3D 体积, 如 (c、d) 所示。在 (a、b) 中, 此类卷的轮廓在 3D MR 图像框中表示。<a href="//ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55798/55798fig20large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
作为一个重要的和额外的产品, 实验产生了一个扩展的多数据集的美国图像, 包括定位和注册数据相对于同一目标的不同 MR 图像。图 21将数据集中的所有3D 图像汇总到相同的 MR 图像中, 从而将它们的边界框显示在一起。
<img alt="Figure 21" class="xfigimg" src="/files/ftp_upload/55798/55798fig21.jpg"> 
图 21: 在 MR 图像参考帧中获取3D 美国数据集.该图显示了3D 的大脑 MR 图像和与压电机械扫掠 (a)、压电线性阵列 (b) 和 CMUT (c) 探针所获得的3D 美国数据集的叠加轮廓。在 (b) 和 (c) 中, 通过容积重建获得了3D 图像。<a href="//ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55798/55798fig21large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
总的来说, 这些结果证明了所描述的协议的有效性, 它允许在同一牛脑的 3D MR 图像 (图 18, 19) 的参考帧中获取和正确注册2维/3 维美国图像,并重建从2D 美国图像在手绘模式获得的卷 (数字 19d, 20)。
使用所描述的软件工具, 专家可以在2D 和3D 美国生物标本的图像中探索最重要的特征。与其他美国探头相比, CMUT 探针成像性能的定性评估的重要例子 (参见图 18、图 19和图 20) 和参考目标 MR 图像 (见图 18)。通过人类专家或其他软件技术的应用, 如数字的、fine-tuned 的我们-MR 3D 图像的注册, 可以对图像数据集进行进一步的复杂分析。这些软件技术将在未来的工作中得到解决。

Watch this Scientific Journal Video about An Experimental Protocol for Assessing the Performance of New Ultrasound Probes Based on CMUT Technology in Application to Brain Imaging at JoVE.com

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

基于电容式微机械超声换能器 (CMUT) 技术的新型超声探头的研制需要对成像能力进行早期逼真的评估。我们描述了一个可重复的实验协议, 美国图像采集和与磁共振图像的比较, 使用一个前体内牛脑作为成像靶。

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 ...

an-experimental-protocol-for-assessing-performance-new-ultrasound

Universidad San Sebastian

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SCIENTISTS IN ACTION

10.4K 次观看.  University of Pavia. 基于电容式微机械超声换能器 (CMUT) 技术的新型超声探头的研制需要对成像能力进行早期逼真的评估。我们描述了一个可重复的实验协议, 美国图像采集和与磁共振图像的比较, 使用一个前体内牛脑作为成像靶。

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

Functional Neuroimaging Using Ultrasonic Blood-brain Barrier Disruption and Manganese-enhanced MRI

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains technically challenging. One approach, Activation-Induced Manganese-enhanced MRI (AIM MRI), has been used successfully to map neuronal activity in rodents 1-5. In AIM MRI, Mn2+ acts a calcium analog and accumulates in depolarized neurons 6,7. Because Mn2+ shortens the T1 tissue property, regions of elevated neuronal activity will enhance in MRI. Furthermore, Mn2+ clears slowly from the activated regions; therefore, stimulation can be performed outside the magnet prior to imaging, enabling greater experimental flexibility. However, because Mn2+ does not readily cross the blood-brain barrier (BBB), the need to open the BBB has limited the use of AIM MRI, especially in mice.
One tool for opening the BBB is ultrasound. Though potentially damaging, if ultrasound is administered in combination with gas-filled microbubbles (i.e., ultrasound contrast agents), the acoustic pressure required for BBB opening is considerably lower. This combination of ultrasound and microbubbles can be used to reliably open the BBB without causing tissue damage 8-11.
Here, a method is presented for performing AIM MRI by using microbubbles and ultrasound to open the BBB. After an intravenous injection of perflutren microbubbles, an unfocused pulsed ultrasound beam is applied to the shaved mouse head for 3 minutes. For simplicity, we refer to this technique of BBB Opening with Microbubbles and UltraSound as BOMUS 12. Using BOMUS to open the BBB throughout both cerebral hemispheres, manganese is administered to the whole mouse brain. After experimental stimulation of the lightly sedated mice, AIM MRI is used to map the neuronal response.
To demonstrate this approach, herein BOMUS and AIM MRI are used to map unilateral mechanical stimulation of the vibrissae in lightly sedated mice 13. Because BOMUS can open the BBB throughout both hemispheres, the unstimulated side of the brain is used to control for nonspecific background stimulation. The resultant 3D activation map agrees well with published representations of the vibrissae regions of the barrel field cortex 14. The ultrasonic opening of the BBB is fast, noninvasive, and reversible; and thus this approach is suitable for high-throughput and/or longitudinal studies in awake mice.

A technique is described for broadly opening the blood-brain barrier in the mouse using microbubbles and ultrasound. Using this technique, manganese can be administered to the mouse brain. Because manganese is an MRI contrast agent that accumulates in depolarized neurons, this approach enables imaging of neuronal activity.

使用超声波血脑屏障破坏和锰增强MRI的脑功能成像

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains ...

科研

神经科学

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 ...

生物工程

State of the Art Cranial Ultrasound Imaging in Neonates

Cranial ultrasound (CUS) is a reputable tool for brain imaging in critically ill neonates. It is safe, relatively cheap and easy to use, even when a patient is unstable. In addition it is radiation-free and allows serial imaging. CUS possibilities have steadily expanded. However, in many neonatal intensive care units, these possibilities are not optimally used. We present a comprehensive approach for neonatal CUS, focusing on optimal settings, different probes, multiple acoustic windows and Doppler techniques. This approach is suited for both routine clinical practice and research purposes. In a live demonstration, we show how this technique is performed in the neonatal intensive care unit. Using optimal settings and probes allows for better imaging quality and improves the diagnostic value of CUS in experienced hands. Traditionally, images are obtained through the anterior fontanel. Use of supplemental acoustic windows (lambdoid, mastoid, and lateral fontanels) improves detection of brain injury. Adding Doppler studies allows screening of patency of large intracranial arteries and veins. Flow velocities and indices can be obtained. Doppler CUS offers the possibility of detecting cerebral sinovenous thrombosis at an early stage, creating a window for therapeutic intervention prior to thrombosis-induced tissue damage. Equipment, data storage and safety aspects are also addressed.

Cranial ultrasound (CUS) is a valuable tool for brain imaging in critically ill neonates. This video shows a comprehensive approach for neonatal (Doppler) CUS for both clinical and research purposes, including a bedside demonstration of the technique.

艺术颅超声成像在新生儿的状态

Cranial ultrasound (CUS) is a reputable tool for brain imaging in critically ill neonates. It is safe, relatively cheap and easy to use, even when a ...

医学

Non-invasive Parenchymal, Vascular and Metabolic High-frequency Ultrasound and Photoacoustic Rat Deep Brain Imaging

Photoacoustics and high frequency ultrasound stands out as powerful tools for neurobiological applications enabling high-resolution imaging on the central nervous system of small animals. However, transdermal and transcranial neuroimaging is frequently affected by low sensitivity, image aberrations and loss of space resolution, requiring scalp or even skull removal before imaging. To overcome this challenge, a new protocol is presented to gain significant insights in brain hemodynamics by photoacoustic and high-frequency ultrasounds imaging with the animal skin and skull intact. The procedure relies on the passage of ultrasound (US) waves and laser directly through the fissures that are naturally present on the animal cranium. By juxtaposing the imaging transducer device exactly in correspondence to these selected areas where the skull has a reduced thickness or is totally absent, one can acquire high quality deep images and explore internal brain regions that are usually difficult to anatomically or functionally describe without an invasive approach. By applying this experimental procedure, significant data can be collected in both sonic and optoacoustic modalities, enabling to image the parenchymal and the vascular anatomy far below the head surface. Deep brain features such as parenchymal convolutions and fissures separating the lobes were clearly visible. Moreover, the configuration of large and small blood vessels was imaged at several millimeters of depth, and precise information were collected about blood fluxes, vascular stream velocities and the hemoglobin chemical state. This repertoire of data could be crucial in several research contests, ranging from brain vascular disease studies to experimental techniques involving the systemic administration of exogenous chemicals or other objects endowed with imaging contrast enhancement properties. In conclusion, thanks to the presented protocol, the US and PA techniques become an attractive noninvasive performance-competitive means for cortical and internal brain imaging, retaining a significant potential in many neurologic fields.

The present work describes a new protocol to perform non-invasive high-frequency ultrasound and photoacoustic based imaging on rat brain, to efficiently visualize deep subcortical regions and their vascular patterns by directing signals on skull foramina naturally present on animal cranium.

非侵入性的脑实质，血管和代谢高频超声和光声大鼠深脑成像

Photoacoustics and high frequency ultrasound stands out as powerful tools for neurobiological applications enabling high-resolution imaging on the central ...

Real-Time Intravital Multiphoton Microscopy to Visualize Focused Ultrasound and Microbubble Treatments to Increase Blood-Brain Barrier Permeability

The blood-brain barrier (BBB) is a key challenge for the successful delivery of drugs to the brain. Ultrasound exposure in the presence of microbubbles has emerged as an effective method to transiently and locally increase the permeability of the BBB, facilitating para- and transcellular transport of drugs across the BBB. Imaging the vasculature during ultrasound-microbubble treatment will provide valuable and novel insights on the mechanisms and dynamics of ultrasound-microbubble treatments in the brain.
Here, we present an experimental procedure for intravital multiphoton microscopy using a cranial window aligned with a ring transducer and a 20x objective lens. This set-up enables high spatial and temporal resolution imaging of the brain during ultrasound-microbubble treatments. Optical access to the brain is obtained via an open-skull cranial window. Briefly, a 3-4 mm diameter piece of the skull is removed, and the exposed area of the brain is sealed with a glass coverslip. A 0.82 MHz ring transducer, which is attached to a second glass coverslip, is mounted on top. Agarose (1% w/v) is used between the coverslip of the transducer and the coverslip covering the cranial window to prevent air bubbles, which impede ultrasound propagation. When sterile surgery procedures and anti-inflammatory measures are taken, ultrasound-microbubble treatments and imaging sessions can be performed repeatedly over several weeks. Fluorescent dextran conjugates are injected intravenously to visualize the vasculature and quantify ultrasound-microbubble induced effects (e.g., leakage kinetics, vascular changes). This paper describes the cranial window placement, ring transducer placement, imaging procedure, common troubleshooting steps, as well as advantages and limitations of the method.

This protocol describes the surgical and technical procedures that enable real-time in vivo multiphoton fluorescence imaging of the rodent brain during focused ultrasound and microbubble treatments to increase blood-brain barrier permeability.

实时活体内多光子显微镜可可视化聚焦超声和微气泡处理，以提高血脑屏障通透性

The blood-brain barrier (BBB) is a key challenge for the successful delivery of drugs to the brain. Ultrasound exposure in the presence of microbubbles ...

A Bedside, Single Burr Hole Approach to Multimodality Monitoring in Severe Brain Injury

Intracranial pressure (ICP) monitoring is a cornerstone of the intensive care management of patients with severe acute brain injuries, including traumatic brain injury. While elevations in ICP are common, data regarding the measurement and treatment of these ICP elevations are conflicting. There is increasing recognition that changes in the balance between supply and demand of brain tissue are critically important and therefore the measurement of multiple modalities is required. Approaches are not standard, and therefore this article provides a description of a bedside, single burr hole approach to multimodality monitoring that allows the passage of probes designed to measure not only ICP but brain tissue oxygen, blood flow, and intracranial electroencephalography. Patient selection criteria, operative procedures, and practical considerations for securing probes during critical care are described. This method is readily performed, safe, secure, and flexible for the adoption of a variety of multimodality monitoring approaches aimed at detecting or preventing secondary brain injuries.

A method of recording multimodality monitoring signals in patients with severe brain injuries using a bedside, single burr hole technique is described.

一种用于重型颅脑损伤多模态监测的床边单毛刺孔方法

Intracranial pressure (ICP) monitoring is a cornerstone of the intensive care management of patients with severe acute brain injuries, including traumatic ...

Focused Ultrasound Induced Blood-Brain Barrier Opening for Targeting Brain Structures and Evaluating Chemogenetic Neuromodulation

Acoustically Targeted Chemogenetics (ATAC) allows for the noninvasive control of specific neural circuits. ATAC achieves such control through a combination of focused ultrasound (FUS) induced blood-brain barrier opening (FUS-BBBO), gene delivery with adeno-associated viral (AAV) vectors, and activation of cellular signaling with engineered, chemogenetic, protein receptors and their cognate ligands. With ATAC, it is possible to transduce both large and small brain regions with millimeter precision using a single noninvasive ultrasound application. This transduction can later allow for a long-term, noninvasive, device-free neuromodulation in freely moving animals using a drug. Since FUS-BBBO, AAVs, and chemogenetics have been used in multiple animals, ATAC should also be scalable for the use in other animal species. This paper expands upon a previously published protocol and outlines how to optimize the gene delivery with FUS-BBBO to small brain regions with MRI-guidance but without a need for a complicated MRI-compatible FUS device. The protocol, also, describes the design of mouse targeting and restraint components that can be 3D-printed by any lab and can be easily modified for different species or custom equipment. To aid reproducibility, the protocol describes in detail how the microbubbles, AAVs, and venipuncture were used in ATAC development. Finally, an example data is shown to guide the preliminary investigations of studies utilizing ATAC.

This protocol delineates steps necessary for the gene delivery through focused ultrasound blood brain barrier (BBB) opening, evaluation of the resulting gene expression, and measurement of neuromodulation activity of chemogenetic receptors through histological tests.

聚焦超声诱导血脑屏障打开，用于靶向大脑结构和评估化学遗传神经调控

Acoustically Targeted Chemogenetics (ATAC) allows for the noninvasive control of specific neural circuits. ATAC achieves such control through a ...

Whole-Brain 3D Activation and Functional Connectivity Mapping in Mice using Transcranial Functional Ultrasound Imaging

Functional ultrasound (fUS) imaging is a novel brain imaging modality that relies on the high-sensitivity measure of the cerebral blood volume achieved by ultrafast doppler angiography. As brain perfusion is strongly linked to local neuronal activity, this technique allows the whole-brain 3D mapping of task-induced regional activation as well as resting-state functional connectivity, non-invasively, with unmatched spatio-temporal resolution and operational simplicity. In comparison with fMRI (functional magnetic resonance imaging), a main advantage of fUS imaging consists in enabling a complete compatibility with awake and behaving animal experiments. Moreover, fMRI brain mapping in mice, the most used preclinical model in Neuroscience, remains technically challenging due to the small size of the brain and the difficulty to maintain stable physiological conditions. Here we present a simple, reliable and robust protocol for whole-brain fUS imaging in anesthetized and awake mice using an off-the-shelf commercial fUS system with a motorized linear transducer, yielding significant cortical activation following sensory stimulation as well as reproducible 3D functional connectivity pattern for network identification.

This protocol describes the quantification of volumetric cerebral hemodynamic variations in the mouse brain using functional ultrasound (fUS). Procedures for 3D functional activation map following sensory stimulation as well as resting-state functional connectivity are provided as illustrative examples, in anesthetized and awake mice.

使用经颅功能超声成像的小鼠全脑3D激活和功能连接映射

Functional ultrasound (fUS) imaging is a novel brain imaging modality that relies on the high-sensitivity measure of the cerebral blood volume achieved by ...

高精度 TUS：使用声学和热仿真进行规划和执行

Pipeline for Planning and Execution of Transcranial Ultrasound Neuromodulation Experiments in Humans

Transcranial ultrasound stimulation (TUS) is an emerging non-invasive neuromodulation technique capable of manipulating both cortical and subcortical structures with high precision. Conducting experiments involving humans necessitates careful planning of acoustic and thermal simulations. This planning is essential to adjust for bone interference with the ultrasound beam's shape and trajectory and to ensure TUS parameters meet safety requirements. T1- and T2-weighted, along with zero-time echo (ZTE) magnetic resonance imaging (MRI) scans with 1 mm isotropic resolution, are acquired (alternatively computed tomography x-ray (CT) scans) for skull reconstruction and simulations. Target and trajectory mapping are performed using a neuronavigational platform. SimNIBS is used for the initial segmentation of the skull, skin, and brain tissues. Simulation of TUS is carried over with the BabelBrain tool, which uses the ZTE scan to produce synthetic CT images of the skull to be converted into acoustic properties. We use a phased array ultrasound transducer with electrical steering capabilities. Z-steering is adjusted to ensure that the target depth is reached. Other transducer configurations are also supported in the planning tool. Thermal simulations are run to ensure temperature and mechanical index requirements are within the acoustic guidelines for TUS in human subjects as recommended by the FDA. During TUS delivery sessions, a mechanical arm assists in the movement of the transducer to the required location using a frameless stereotactic localization system.

作者聚焦：推进人脑调节 - 经颅超声刺激实验的优化方案

Transcranial ultrasound stimulation (TUS) is an emerging non-invasive neuromodulation technique that requires careful planning of acoustic and thermal simulations. The methodology describes an image processing and ultrasound simulation pipeline for efficient, user-friendly, streamlined planning for human TUS experimentation.

用于规划和执行人体经颅超声神经调控实验的管道

Transcranial ultrasound stimulation (TUS) is an emerging non-invasive neuromodulation technique capable of manipulating both cortical and subcortical ...

基于 CMUT 技术的新型超声探头在脑成像中的应用研究

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一种用于重型颅脑损伤多模态监测的床边单毛刺孔方法

聚焦超声诱导血脑屏障打开，用于靶向大脑结构和评估化学遗传神经调控

使用经颅功能超声成像的小鼠全脑3D激活和功能连接映射

用于规划和执行人体经颅超声神经调控实验的管道

用于规划和执行人体经颅超声神经调控实验的管道