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。

<ol>
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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...

<table border="0" cellpadding="0" cellspacing="0" width="925">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">ULA-OP</td>
			<td style="width:167px;">University of Florence</td>
			<td style="width:104px;">N/A</td>
			<td style="width:415px;">Ultrasound imaging research system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">3D imaging piezeoelectric probe</td>
			<td style="width:167px;">Esaote s.p.a.</td>
			<td style="width:104px;">9600195000</td>
			<td>Mechanically-swept 3D ultrasound probe, model BL-433</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Linear-array piezoelectric probe</td>
			<td style="width:167px;">Esaote s.p.a.</td>
			<td style="width:104px;">122001100</td>
			<td>Ultrasound linear array probe, model LA-533</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">CMUT probe</td>
			<td style="width:167px;">University Roma Tre</td>
			<td style="width:104px;">N/A</td>
			<td>Ultrasound linear array probe based on CMUT technology</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">MAGNETOM Skyra 3T MR scanner</td>
			<td style="width:167px;">Siemens Healthcare</td>
			<td style="width:104px;">N/A</td>
			<td>MR scanner</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Head coil</td>
			<td style="width:167px;">Siemens Healthcare</td>
			<td style="width:104px;">N/A</td>
			<td>32-channel head coil for MR imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">NDI Polaris Vicra</td>
			<td style="width:167px;">NDI Medical</td>
			<td style="width:104px;">8700335001</td>
			<td>Optical motion tracking system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Pointer tool</td>
			<td style="width:167px;">NDI Medical</td>
			<td style="width:104px;">8700340</td>
			<td>Passive pointer tool with 4 reflecting markers</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Clamp-equipped tool</td>
			<td style="width:167px;">NDI Medical</td>
			<td style="width:104px;">8700399</td>
			<td>Rigid body with 4 reflecting markers and a clamp to be connected to the US probe handle</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Bovine brain</td>
			<td style="width:167px;">N/A</td>
			<td style="width:104px;">N/A</td>
			<td>Brain of an adult bovine, from food suppliers</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Formalin solution</td>
			<td style="width:167px;">N/A</td>
			<td style="width:104px;">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 height="21">
			<td height="21" style="height:21px;width:240px;">Plastic container for anatomical parts</td>
			<td style="width:167px;">N/A</td>
			<td style="width:104px;">N/A</td>
			<td>Cilindrical plastic container with lid</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Glass spheres</td>
			<td style="width:167px;">N/A</td>
			<td style="width:104px;">N/A</td>
			<td>3mm diameter spheres of Flint glass</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Agar</td>
			<td style="width:167px;">N/A</td>
			<td style="width:104px;">N/A</td>
			<td>30 g, for phantom preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Glycerine</td>
			<td style="width:167px;">AEFFE Farmaceutici</td>
			<td style="width:104px;">A908005248</td>
			<td>100 g, for phantom preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Distilled water</td>
			<td style="width:167px;">Solbat Gaysol</td>
			<td style="width:104px;">8027391000015</td>
			<td>870 g, for phantom preparation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Beaker</td>
			<td style="width:167px;">N/A</td>
			<td style="width:104px;">N/A</td>
			<td>Beaker used for the diluition of glycerine and agar in distilled water</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Lysoform</td>
			<td style="width:167px;">Lever</td>
			<td style="width:104px;">8000680500014</td>
			<td>A benzalkonium chloride and water solution was used for the agar phantom preservation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Polystyrene mannequin head</td>
			<td style="width:167px;">N/A</td>
			<td style="width:104px;">N/A</td>
			<td>Polyestirene model which was cutted and used to design the configuration of spheres&#39;patterns</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Green tissue marking dye for histology</td>
			<td style="width:167px;">N/A</td>
			<td style="width:104px;">N/A</td>
			<td>Colour used to mark the glass spheres&#39; positions on the bovine brain surface</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Yellow enamel</td>
			<td style="width:167px;">N/A</td>
			<td style="width:104px;">N/A</td>
			<td>Enamel used to colour the glass spheres implanted in the agar phantom</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Water tank</td>
			<td style="width:167px;">N/A</td>
			<td style="width:104px;">N/A</td>
			<td>50x50x30 cm plastic tank filled degassed water up to a 15cm height&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Mechanical arm</td>
			<td style="width:167px;">Esaote s.p.a.</td>
			<td style="width:104px;">N/A</td>
			<td>Mechanical arm clamped to the water tank border and used to held the probe in fixed positions</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Plate of synthetic resin</td>
			<td style="width:167px;">N/A</td>
			<td style="width:104px;">N/A</td>
			<td>Plate used as a support for the bovine brain positioning in the water tank</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Sewing threads</td>
			<td style="width:167px;">N/A</td>
			<td style="width:104px;">N/A</td>
			<td>Sewing thread segments used to immobilize the brain on the resin plate</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Adhesive tape</td>
			<td style="width:167px;">N/A</td>
			<td style="width:104px;">N/A</td>
			<td>Adhesive tape used to fix the sewing thread extremities onto the resin plate</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Plastic food container</td>
			<td style="width:167px;">N/A</td>
			<td style="width:104px;">N/A</td>
			<td>Sealed food container used for the agar phantom</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Notebook</td>
			<td style="width:167px;">Lenovo</td>
			<td style="width:104px;">Z50-70</td>
			<td>Lenovo&#160; Z50-70, Intel(R) Core i7-4510U @ 2.0 GHz, 8 GB RAM</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Workstation</td>
			<td style="width:167px;">Dell Inc.</td>
			<td style="width:104px;">T5810</td>
			<td>Intel(R) Xeon(R) CPU E3-1240v3 @ 3.40 GHz, 16 GB RAM</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Matlab</td>
			<td style="width:167px;">The MathWorks</td>
			<td style="width:104px;">R2013a</td>
			<td>Software tool, used for space transformation computation and 3D reconstruction from image planes</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">Paraview</td>
			<td style="width:167px;">Kitware Inc.</td>
			<td style="width:104px;">v. 4.4.1</td>
			<td>Open-source software for 3D image processing and visualization</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">NDI Toolbox - ToolTracker Utility</td>
			<td style="width:167px;">NDI Medical</td>
			<td style="width:104px;">v. 4.007.007</td>
			<td>Software for marker position visualization and tracking in the NDI Polaris Vicra measurement volume</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">C++ data-logging software</td>
			<td style="width:167px;">NDI Medical</td>
			<td style="width:104px;">v. 4.007.007</td>
			<td>Software for marker position recording on a text log file</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:240px;">ULA-OP software&#160;</td>
			<td style="width:167px;">University of Florence</td>
			<td style="width:104px;">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...

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

Research

Education

JoVE Core

JoVE Journal

Electrical Engineering

Bioengineering

Unit 1

Diagrams and Signal Flow Graphs

SCIENTISTS IN ACTION

10.3K 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

High-throughput, Automated Extraction of DNA and RNA from Clinical Samples using TruTip Technology on Common Liquid Handling Robots

TruTip is a simple nucleic acid extraction technology whereby a porous, monolithic binding matrix is inserted into a pipette tip. The geometry of the monolith can be adapted for specific pipette tips ranging in volume from 1.0 to 5.0 ml. The large porosity of the monolith enables viscous or complex samples to readily pass through it with minimal fluidic backpressure. Bi-directional flow maximizes residence time between the monolith and sample, and enables large sample volumes to be processed within a single TruTip. The fundamental steps, irrespective of sample volume or TruTip geometry, include cell lysis, nucleic acid binding to the inner pores of the TruTip monolith, washing away unbound sample components and lysis buffers, and eluting purified and concentrated nucleic acids into an appropriate buffer. The attributes and adaptability of TruTip are demonstrated in three automated clinical sample processing protocols using an Eppendorf epMotion 5070, Hamilton STAR and STARplus liquid handling robots, including RNA isolation from nasopharyngeal aspirate, genomic DNA isolation from whole blood, and fetal DNA extraction and enrichment from large volumes of maternal plasma (respectively).

TruTip is a simple nucleic acid extraction technology whereby a porous, monolithic binding matrix is inserted into a pipette tip. Consequently, the sample preparation format is compatible with most liquid handling instruments, and can be used for many medium to high-throughput clinical applications and sample types.

高通量，自动化的DNA和RNA的提取从临床样品使用TruTip技术常见的液体处理机器人

TruTip is a simple nucleic acid extraction  technology whereby a porous, monolithic binding matrix is inserted into a  pipette tip. The geometry of the ...

科研

生物工程

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 ...

In Situ Mapping of the Mechanical Properties of Biofilms by Particle-tracking Microrheology

Bacterial cells are able to form surface-attached biofilm communities known as biofilms by encasing themselves in extracellular polymeric substances (EPS). The EPS serves as a physical and protective scaffold that houses the bacterial cells and consists of a variety of materials that includes proteins, exopolysaccharides and DNA. The composition of the EPS may change, which remodels the mechanic properties of the biofilm to further develop or support alternative biofilm structures, such as streamers, as a response to environmental cues. Despite this, there are little quantitative descriptions on how EPS components contribute to the mechanical properties and function of biofilms. Rheology, the study of the flow of matter, is of particular relevance to biofilms as many biofilms grow in flow conditions and are constantly exposed to shear stress. It also provides measurement and insight on the spreading of the biofilm on a surface. Here, particle-tracking microrheology is used to examine the viscoelasticity and effective crosslinking roles of different matrix components in various parts of the biofilm during development. This approach allows researchers to measure mechanic properties of biofilms at the micro-scale, which might provide useful information for controlling and engineering biofilms.

In Situ Mapping of the Mechanical Properties of Biofilms by Particle-tracking Microrheology

Particle-tracking microrheology investigates the viscoelasticity of materials. Here, the technique is used to determine the viscoelasticity, creep compliance and effective crosslinking roles of different matrix components of a bacterial biofilm. The matrix consists of polymeric substances secreted by the bacteria and its components determine biofilm structure and mechanical properties.

在生物膜的力学性能原位制图粒子追踪Microrheology

Bacterial cells are able to form surface-attached biofilm communities known as biofilms by encasing themselves in extracellular polymeric substances ...

An Experimental and Finite Element Protocol to Investigate the Transport of Neutral and Charged Solutes across Articular Cartilage

Osteoarthritis (OA) is a debilitating disease that is associated with degeneration of articular cartilage and subchondral bone. Degeneration of articular cartilage impairs its load-bearing function substantially as it experiences tremendous chemical degradation, i.e. proteoglycan loss and collagen fibril disruption. One promising way to investigate chemical damage mechanisms during OA is to expose the cartilage specimens to an external solute and monitor the diffusion of the molecules. The degree of cartilage damage (i.e. concentration and configuration of essential macromolecules) is associated with collisional energy loss of external solutes while moving across articular cartilage creates different diffusion characteristics compared to healthy cartilage. In this study, we introduce a protocol, which consists of several steps and is based on previously developed experimental micro-Computed Tomography (micro-CT) and finite element modeling. The transport of charged and uncharged iodinated molecules is first recorded using micro-CT, which is followed by applying biphasic-solute and multiphasic finite element models to obtain diffusion coefficients and fixed charge densities across cartilage zones.

We propose a protocol to investigate the transport of charged and uncharged molecules across articular cartilage with the aid of recently developed experimental and numerical methods.

实验和有限元议定书调查中性和带电溶质的跨关节软骨运输

Osteoarthritis (OA) is a debilitating disease that is associated with degeneration of articular cartilage and subchondral bone. Degeneration of articular ...

A High-performance Compact Photoacoustic Tomography System for In Vivo Small-animal Brain Imaging

In vivo small-animal imaging has an important role to play in preclinical studies. Photoacoustic tomography (PAT) is an emerging hybrid imaging modality that shows great potential for both preclinical and clinical applications. Conventional optical parametric oscillator-based PAT (OPO-PAT) systems are bulky and expensive and cannot provide high-speed imaging. Recently, pulsed-laser diodes (PLDs) have been successfully demonstrated as an alternative excitation source for PAT. Pulsed-laser diode PAT (PLD-PAT) has been successfully demonstrated for high-speed imaging on photoacoustic phantoms and biological tissues. This work provides a visualized experimental protocol for in vivo brain imaging using PLD-PAT. The protocol includes the compact PLD-PAT system configuration and its description, animal preparation for brain imaging, and a typical experimental procedure for 2D cross-sectional rat brain imaging. The PLD-PAT system is compact and cost-effective and can provide high-speed, high-quality imaging. Brain images collected in vivo at various scan speeds are presented.

A High-performance Compact Photoacoustic Tomography System for In Vivo Small-animal Brain Imaging

A compact pulsed-laser diode-based photoacoustic tomography (PLD-PAT) system for high-speed in vivo brain imaging in small animals is demonstrated.

高性能紧凑型光声层析成像系统<em&gt;体内</em&gt;小动物脑成像

In vivo small-animal imaging has an important role to play in preclinical studies. Photoacoustic tomography (PAT) is an emerging hybrid imaging modality ...

Fabrication and Validation of an Organ-on-chip System with Integrated Electrodes to Directly Quantify Transendothelial Electrical Resistance

Organs-on-chips, in vitro models involving the culture of (human) tissues inside microfluidic devices, are rapidly emerging and promise to provide useful research tools for studying human health and disease. To characterize the barrier function of cell layers cultured inside organ-on-chip devices, often transendothelial or transepithelial electrical resistance (TEER) is measured. To this end, electrodes are usually integrated into the chip by micromachining methods to provide more stable measurements than is achieved with manual insertion of electrodes into the inlets of the chip. However, these electrodes frequently hamper visual inspection of the studied cell layer or require expensive cleanroom processes for fabrication. To overcome these limitations, the device described here contains four easily integrated electrodes that are placed and fixed outside of the culture area, making visual inspection possible. Using these four electrodes the resistance of six measurement paths can be quantified, from which the TEER can be directly isolated, independent of the resistance of culture medium-filled microchannels. The blood-brain barrier was replicated in this device and its TEER was monitored to show the device applicability. This chip, the integrated electrodes and the TEER determination method are generally applicable in organs-on-chips, both to mimic other organs or to be incorporated into existing organ-on-chip systems.

This publication describes the fabrication of an organ-on-chip device with integrated electrodes for direct quantification of transendothelial electrical resistance (TEER). For validation, the blood-brain barrier was mimicked inside this microfluidic device and its barrier function was monitored. The presented methods for electrode integration and direct TEER quantification are generally applicable.

用集成电极直接量化 Transendothelial 电阻的 Organ-on-chip 系统的制作与验证

Organs-on-chips, in vitro models involving the culture of (human) tissues inside microfluidic devices, are rapidly emerging and promise to provide useful ...

Agarose-based Tissue Mimicking Optical Phantoms for Diffuse Reflectance Spectroscopy

This protocol describes how to make agarose-based tissue-mimicking phantoms and demonstrates how to determine their optical properties using a conventional optical system with an integrating sphere. Measuring systems for the acquisition of the diffuse reflectance and total transmittance spectra are constructed with a broadband white light source, a light guide, an achromatic lens, an integrating sphere, a sample holder, an optical fiber probe, and a multi-channel spectrometer. An acrylic mold consisting of two rectangular acrylic pieces and a U-shaped acrylic piece is constructed to create an epidermal phantom and a dermal phantom with whole blood. The application of a sodium dithionite (Na2S2O4) solution to the dermal phantom enables the researcher to deoxygenate hemoglobin in red blood cells distributed in the dermal phantom. The inverse Monte Carlo simulation with the diffuse reflectance and total transmittance spectra measured by a spectrometer with an integrating sphere is performed to determine the absorption coefficient spectrum &#181;a(&#955;) and the reduced scattering coefficient spectrum &#181;s&#39;(&#955;) of each layer phantom. A two-layered phantom mimicking the diffuse reflectance of human skin tissue is also demonstrated by piling up the epidermal phantom on the dermal phantom.

Here, we demonstrate how agarose-based tissue-mimicking optical phantoms are made and how their optical properties are determined using a conventional optical system with an integrating sphere.

基于琼脂糖组织模拟光学幽灵的漫反射光谱研究

This protocol describes how to make agarose-based tissue-mimicking phantoms and demonstrates how to determine their optical properties using a ...

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver engineering, including in vivo organ decellularization followed by repopulation, has emerged as a promising approach over ex vivo liver engineering. However, postoperative survival was not achieved. The aim of this study is to develop a novel surgical technique of in vivo selective liver lobe perfusion in rats as a prerequisite for in vivo liver engineering. We generate a circuit bypass only through the left lateral lobe. Then, the left lateral lobe is perfused with heparinized saline. The experiment is performed with 4 groups (n = 3 rats per group) based on different perfusion times of 20 min, 2 h, 3 h, and 4 h. Survival, as well as the macroscopically visible change of color and the histologically determined absence of blood cells in the portal triad and the sinusoids, is taken as an indicator for a successful model establishment. After selective perfusion of the left lateral lobe, we observe that the left lateral lobe, indeed, turned from red to faint yellow. In a histological assessment, no blood cells are visible in the branch of the portal vein, the central vein, and the sinusoids. The left lateral lobe turns red after reopening the blocked vessels. 12/12 rats survived the procedure for more than one week. We are the first to report a surgical model for in vivo single liver lobe perfusion with a long survival period of more than one week. In contrast to the previously published report, the most important advantage of the technique presented here is that perfusion of 70% of the liver is maintained throughout the whole procedure. The establishment of this technique provides a foundation for in vivo partial liver engineering in rats, including decellularization and recellularization.

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

We establish a novel surgical technique for an in vivo single liver lobe perfusion model in rat as a prerequisite for further studying in vivo partial liver engineering in the future.

大鼠体内部分肝工程的外科技术基础

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver ...

A New Ex Vivo Model for the Evaluation of Endoscopic Submucosal Injection Material Performance

Increasing the performance of submucosal injection materials (SIMs) is important for endoscopic therapy of early gastrointestinal cancer. It is essential to establish an ex vivo model that can evaluate SIM performance accurately, for developing high-performance SIMs. In our previous study, we developed a new ex vivo model that can be used to evaluate the performance of various SIMs in detail by applying constant tension to the specimen&#39;s ends. We also confirmed that the proposed new ex vivo model allows accurate submucosal elevation height (SEH) measurement under uniform conditions and detailed comparisons of the performances of various types of SIMs. Here, we describe the new ex vivo model and explain the detailed setup methodology of this model. Since all parts of the new model were easy to obtain, the setup of the new model could be completed quickly. SEH of various SIMs could be measured more accurately by using the new model. The critical factor that determines SIM performance can be identified using the new model. SIM development speed will drastically increase after the factor has been identified.

A New Ex Vivo Model for the Evaluation of Endoscopic Submucosal Injection Material Performance

We developed a new ex vivo model that applies constant tension to the porcine gastric specimen. This development made it possible to evaluate the performance (the height and duration of the submucosal elevation) of various SIMs accurately.&#160; The detailed setup methodology of this new model is explained.

内镜下黏膜下注射材料性能评价的一种新的体内模型

Increasing the performance of submucosal injection materials (SIMs) is important for endoscopic therapy of early gastrointestinal cancer. It is essential ...

An Open Source Technology Platform to Manufacture Hydrogel-Based 3D Culture Models in an Automated and Standardized Fashion

Current mixing steps of viscous materials rely on repetitive and time-consuming tasks which are performed mainly manually in a low throughput mode. These issues represent drawbacks in workflows that can ultimately result in irreproducibility of research findings. Manual-based workflows are further limiting the advancement and widespread adoption of viscous materials, such as hydrogels used for biomedical applications. These challenges can be overcome by using automated workflows with standardized mixing processes to increase reproducibility. In this study, we present step-by-step instructions to use an open source protocol designer, to operate an open source workstation, and to identify reproducible mixtures. Specifically, the open source protocol designer guides the user through the experimental parameter selection and generates a ready-to-use protocol code to operate the workstation. This workstation is optimized for pipetting of viscous materials to enable automated and highly reliable handling by the integration of temperature docks for thermoresponsive materials, positive displacement pipettes for viscous materials, and an optional tip touch dock to remove excess material from the pipette tip. The validation and verification of mixtures are performed by a fast and inexpensive absorbance measurement of Orange G. This protocol presents results to obtain 80% (v/v) glycerol mixtures, a dilution series for gelatin methacryloyl (GelMA), and double network hydrogels of 5% (w/v) GelMA and 2% (w/v) alginate. A troubleshooting guide is included to support users with protocol adoption. The described workflow can be broadly applied to a number of viscous materials to generate user-defined concentrations in an automated fashion.

This protocol serves as a comprehensive tutorial for standardized and reproducible mixing of viscous materials with a novel open source automation technology. Detailed instructions are provided on the operation of a newly developed open source workstation, the usage of an open source protocol designer, and the validation and verification to identify reproducible mixtures.

以自动化和标准化方式制造基于水凝胶的3D培养模型的开源技术平台

Current mixing steps of viscous materials rely on repetitive and time-consuming tasks which are performed mainly manually in a low throughput mode. These ...