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1。 PEDOT：PSS溶液的制备<ol><li>至50ml的PEDOT：PSS，添加乙二醇（增加导电性）以1:4的体积比（乙烯乙二醇对PEDOT：PSS）中，十二烷基苯磺酸（DBSA）0.5微升/毫升作为表面活性剂，和10 mg /毫升3 - 环氧丙氧基丙基（GOPS）作为交联剂，以促进导电性聚合物的玻璃载片的附着性。 </li></ol> 2。 OECT制作（图3） <ol><li>通过剥离光刻定义热蒸发金源极和漏极接点： <ol><li>旋转涂覆光刻胶在一个普通的干净的载玻片上以3000rpm，持续30秒。玻璃尺寸为3英寸×1英寸</li><li>通过光刻定义模式。尺寸可以被改变，但此处列出的尺寸被证明导致最佳的灵敏度。然后使用一个开发者。 </li><li>蒸发5nm的铬和金的100nm时，分别。 </li><li>剥离光致抗蚀剂中的交流一通浴1小时，使与源极和漏极区仅接触面积的衬底。 </li></ol></li><li>该PEDOT的所需长度：PSS信道是1毫米。这是通过使用聚对二甲苯-C（霸-C）的剥离法来图案实现的： <ol><li>加载3.5克霸-C涂层中的设置。均匀地蒸发2微米的聚对二甲苯-C上用Au接触所述基板的顶部。 </li><li>状态的通道，通过光刻： <ol><li>旋涂光致抗蚀剂以3000rpm进行30秒。 </li><li>使用遮罩来照亮通道和紫外线镀金触点20秒，曝光的光刻胶将在开发者变得可溶。 </li><li>使用开发者对光刻胶打开通道面积。 </li></ol></li><li>通过15分钟的血浆步骤（O 2（50 SCCM）和瑞士法郎4（3 SCCM）等离子在160瓦），以打开通道和黄金触点腐蚀霸-C在通道区。 </li></ol></li><li>通过旋涂在PSS混合溶液：存入的PEDOT500 rpm离心45秒。烘烤30秒，在110℃下</li><li>剥离霸-C来揭示PEDOT：下面的基板PSS的通道。该通道应略微重叠与金接触，协议1进行。 </li><li>烘烤的样品1小时，在140℃的大气条件下。 </li></ol> 3。设备组装<ol><li>通过在固化在碱性溶液解决方案的比1:10混合两个解决方案（通常一起提供的套件）使PDMS。在120℃下烤1小时</li><li>通过使用打孔设计良好，切口周边所需面积的正方形。 </li><li>粘上PDMS以及在所述通道的顶部，以产生大约6 mm 2的信道区域。确保源极和漏极触点不包括在内。 </li><li>做一个塑料支撑在它的孔（可以使用支架的细胞培养插入），然后把它粘在PDMS上。待其干燥过夜。 </li><li>测试很适合用笏预填漏呃。 </li><li>焊上锡源极和漏极接触电线。 </li></ol> 4。细胞培养<ol><li>准备细胞培养基如下： <ol><li>在DMEM培养液中，添加1％谷氨酰胺，10％胎牛血清，0.5％青霉素 - 链霉素，和0.1％庆大霉素。 </li><li>使用无菌过滤装置灭菌的细胞培养基。 </li></ol></li><li>在5％CO 2的潮湿气氛保持通道49和68之间的Caco-2细胞在37℃下，在细胞培养介质。 </li><li>使用胰蛋白酶和种子以1.5×10 4个细胞/插入分裂的细胞，每星期一次。 </li><li> 3周以上改变细胞培养基每周两次。 </li></ol> 5。测量与OECT <ol><li>将Ag / AgCl电极导线连接到源表用作栅电极。沉浸前端在细胞培养基中，这是用来作为电解液， 如图1a中 。 </li><li>连接导线从源极和漏极到t他数字源表（双通道设置）。 </li><li>施加矩形脉冲的正电压的栅极和源极之间（V GS）（作为参考电极：接地）。漏极和源极（V DS）之间的负恒定电压被施加和相应电流（I DS）进行测定。 </li><li>使用一台PC正在运行的程序的数据收集。在定制的采集程序进入OECT参数应该如下：V DS =-0.2V，V GS = 0.3 V，V GS时间= 2秒，关闭时间= 28秒。 </li><li>读出的源极-漏极电流（I DS）和栅电流（I GS）。进行测量几分钟，以确保一个稳定的基线信号。 </li></ol> 6。细胞融合与OECT量测注：在实验之前，使细胞层的完整性可以通过测量TER与阻抗谱装置进行验证。每3周大的Caco-2细胞樱雪的TER室温应高于400Ω。厘米2。 <ol><li>在OECT测定关断时间：从电解液中取出栅电极，把细胞培养插管，并更换细胞培养插管内的栅电极。 </li><li>进行与细胞数分钟的基线测量，以确保一个稳定的信号。此基线将被用作用于对应于一个完整的单层（0）的归一化值的计算。 </li></ol> 7。准备和有毒化合物简介<ol><li>准备EGTA的1M溶液在去离子水中，先调节溶液至7.4，用Tris碱将pH。 </li><li>添加EGTA溶液在适当的体积，得到（ 例如在5毫米和100毫米）的所需浓度并考虑到体积。在EGTA应在基底室在关断期间被加入。 </li><li>连续90分钟在协议5衡量。如果测量是北纳克进行在室温下，该细胞的稳定性不会在90分钟后保持不变。 </li><li>在运行结束时，划伤细胞层导致的势垒层和测量15分钟的完全破坏，这一点可以通过去除过滤器并浸渍在媒体中的栅电极来​​完成。此基线将被用作用于对应于一个被破坏单层的归一化值的计算。 </li></ol>

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该项目由欧盟第七框架计划 - 人-2009-RG，居里夫人项目编号：256367（CELLTOX）和欧洲研究委员会ERC-2010-STG建议无258966（IONOSENSE），以及联合授予来自行政法院区域去普罗旺斯阿尔卑斯蔚蓝海岸和CDL制药为ST。我们感谢乔治Malliaras的有关OECT设计和功能有益的讨论。

这种技术提供了一种新的方法将有机电化学晶体管集成与活细胞来衡量屏障组织的完整性。该技术的主要优点是快速性和灵敏度，而且该设备对于阻挡薄纸动态监测的成本低。 这种方法是用活细胞，一个关键点就是一定要使用单层，它代表一个完整的隔离层。阻挡层的参数应的细胞系的表征过程中被定义。因此，至关重要的是不破坏细胞层时，栅电极浸在电解液中对细胞层?...

胃肠上皮屏障的组织，它控制着身体的不同隔室之间的分子通路的一个例子。上皮细胞是由蛋白质所提供物理屏障1对病原体和毒素复合物结合在一起细长的柱状细胞，同时允许水和维持身体所需的营养物质通过。这种选择性是由于上皮细胞的极化，从而产生两种不同的膜结构域：暴露于管腔和锚定在下层组织2,3的细胞的基底侧的细胞的顶侧。紧密连接（TJ）是存在于上皮细胞的顶端部分的蛋白质复合物，并且是一个更大，更复杂称为心尖接头4的一部分。跨越障碍组织离子流可以或者通过跨细胞（通过细胞）或通过细胞旁路（两个相邻小区之间的）途径去。的总和通过两条途径的传输被称为跨上皮电阻。顶交界负责离子和分子通过一个特定的开启和关闭功能，通过跨越障碍5,6监管。这些蛋白质复合物的功能障碍或破坏往往与疾病7-11。此外，许多肠道病原体/毒素是已知的专门针对这种复杂的，由此进入体内并导致腹泻，最有可能作为离子/水流的跨阻挡层12-14块状失调的结果。阻挡薄纸也可通过改变细胞外微环境的改变。钙粘蛋白是细胞 - 细胞粘附的关键蛋白，并参与根尖结的形成。钙是必需的钙粘蛋白的正确构象的结构，并且在细胞外钙的降低已被证明能导致细胞 - 细胞连接的破坏和随后的开口小区15间旁细胞途径。在这项研究中，EGTA（乙二醇 - 双（β-氨基乙醚）-N，N，N&#39;，N&#39;-四乙酸），一个特定的钙离子螯合剂，是用来诱导屏障组织中一个缺口，因为它有已经显示出对细胞旁一个快速和剧烈影响离子流16,17。此钙离子螯合剂被用于对Caco-2细胞线的汇合和分化的单层。中培养细胞培养物的插入，该细胞系是已知的开发在胃肠道的特性而被广泛应用于制药工业，以测试药物18,19的吸收。 方法监测屏障组织的完整性很丰富。这些方法往往光，靠已知的在心尖结20特定蛋白质的免疫荧光染色，或依赖于荧光示踪分子，通常不渗透到阻挡薄纸的定量21,22。然而，无标记的方法（ 即没有荧光团/色）是优选的，因为使用的标签可以产生伪影，并且通常会增加成本和测定时间。电气，无标记的屏障组织的监控，最近成为一个动态监测方法23。例如在电阻抗谱最近的技术进步已经允许市售扫描装置24,25，可以测量跨上皮电阻（TER），穿过细胞层中的离子电导的测定的发展。 有机电子创造了一个独特的机会，通过使用导电聚合物，可以进行电子和离子载体接口电子和生物26,27 28,29的世界。一种新的技术来检测违规使用OECT 30-32屏障组织在最近推出。该设备对现有的技术，我们进行了验证教育署，以评估使用Cellzscope屏障组织的完整性，包括使用荧光黄免疫荧光，透气性试验和阻抗谱。在所有测试的有毒化合物的情况下，OECT被发现具有相同或更好的灵敏度操作，并且具有增加的时间分辨率相比，上述技术。在该装置中，PEDOT：PSS，已被证明是稳定的和生物相容33,34的导电聚合物，用作晶体管通道的活性材料。该OECT组成上的导电聚合物通道的任一侧的漏极和源极电极。这随后被放置在与电解质，从而形成了装置的一个组成部分的接触。栅电极浸在电解液中（ 图1），而当一个正栅极电压施加于栅极，从电解质阳离子被迫进入通道，从而脱掺杂的导电聚合物，并导致在源极-漏极的变化电流。在DEVICE因此，在离子通量由于放大的晶体管微小变化极其敏感。生长在细胞培养插入一个细胞层置于所述栅极电极和所述导电聚合物通道之间。一个完整的细胞层存在下充当屏障的阳离子进入导电聚合物，因此，在一个完整的单层的存在下，漏极电流减小（ 图2：从过渡区域a到b）。在一种有毒化合物的存在下，在阻挡薄纸将逐渐失去其完整性，又让阳离子进入到聚合物膜和增加的漏电流（ 图2：区域c）所示。用这种技术，违反阻挡薄纸在由漏极电流的调制看出，对应于整个单层的磁通的调制。该设备能够测量与实时前所未有的时间分辨率和灵敏度离子通量的微小变化。这项技术西港岛线L是在毒理学作药物测试，诊断疾病或基础研究作为屏障模型可以很容易适应的领域的兴趣。这种方法也将有助于减少动物实验，因为它允许在体外模型的验证，以取代体内试验。

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sensing of barrier tissue disruption with an organic electrochemical transistor

胃肠道是阻挡薄纸，可提供针对病原体和毒素的条目的物理屏障，同时允许必要的离子和分子的推移的一个例子。在此屏障违反可以通过在细胞外钙离子浓度的降低而引起。这减少了钙的浓度会导致在涉及屏障的密封蛋白的构象变化，从而导致增加的细胞旁通量。模仿这种效果的钙离子螯合剂乙二醇 - 双（β-氨基乙基醚）-N，N，N&#39;，N&#39;-四乙酸（EGTA），使用已知的有代表性的胃肠道的细胞单层。不同的方法来检测组织屏障的破坏已经存在，如免疫荧光和透气性测定法。然而，这些方法是耗时且昂贵的，不适合于动态或高通量测量。电子方法测量阻挡薄纸完整性也为跨上皮电阻（TER）的测量存在，但是这些经常是昂贵的和复杂的。快速，便宜，和敏感的方法的发展迫切需要作为屏障组织的完整性是药物发现和病原体/毒素诊断的关键参数。有机电化学晶体管（OECT）集成在一起阻挡薄纸形成细胞已被证明为能够动态监测屏障组织完整性的新设备。该设备能够测量离子通量分钟变化以前所未有时间分辨率和灵敏度，实时，作为屏障组织完整性的指标。这种新方法是基于简单的装置，可以与高通量筛选应用的兼容和制造成本低。

在测量过程中的第一步骤时，漏电流可能略有不同，但在大多数情况下，它应保持稳定（ 图2，部分a）所示。如果该信号是不稳定的时，晶体管应丢弃并更换。这种稳定性检查也确保了设备的电导率任何初始亏损并不会影响后续计量。测量若干分钟后，用细胞形成组织屏障的插入件被放置在通道的顶部。漏极电流应立即降低（ 图2，B部分）。如果漏极电流不减小，这可能...

Watch this Scientific Journal Video about Sensing of Barrier Tissue Disruption with an Organic Electrochemical Transistor at JoVE.com

Sensing of Barrier Tissue Disruption with an Organic Electrochemical Transistor

有机电化学晶体管集成了活细胞，并用于监视整个胃肠道上皮屏障离子流。在本研究中，增加了离子通量，涉及到中断紧密连接的，诱导的钙螯合剂EGTA（乙二醇 - 双（β-氨基乙基醚）-N，N，N&#39;，N&#39;-四乙酸的存在下酸），进行测定。

阻隔组织破坏与有机电化学晶体管传感

The gastrointestinal tract is an example of barrier tissue that provides a physical barrier against entry of pathogens and toxins, while allowing the ...

sensing-barrier-tissue-disruption-with-an-organic-electrochemical

Research

JoVE Journal

Bioengineering

11.6K 次观看.  Ecole Nationale Superieure des Mines. 有机电化学晶体管集成了活细胞，并用于监视整个胃肠道上皮屏障离子流。在本研究中，增加了离子通量，涉及到中断紧密连接的，诱导的钙螯合剂EGTA（乙二醇 - 双（β-氨基乙基醚）-N，N，N'，N'-四乙酸的存在下酸），进行测定。 

视频: Sensing of Barrier Tissue Disruption with an Organic Electrochemical Transistor

Hollow Microneedle-based Sensor for Multiplexed Transdermal Electrochemical Sensing

The development of a minimally invasive multiplexed monitoring system for rapid analysis of biologically-relevant molecules could offer individuals suffering from chronic medical conditions facile assessment of their immediate physiological state. Furthermore, it could serve as a research tool for analysis of complex, multifactorial medical conditions. In order for such a multianalyte sensor to be realized, it must be minimally invasive, sampling of interstitial fluid must occur without pain or harm to the user, and analysis must be rapid as well as selective.
Initially developed for pain-free drug delivery, microneedles have been used to deliver vaccines and pharmacologic agents (e.g., insulin) through the skin.1-2 Since these devices access the interstitial space, microneedles that are integrated with microelectrodes can be used as transdermal electrochemical sensors. Selective detection of glucose, glutamate, lactate, hydrogen peroxide, and ascorbic acid has been demonstrated using integrated microneedle-electrode devices with carbon fibers, modified carbon pastes, and platinum-coated polymer microneedles serving as transducing elements.3-7,8
This microneedle sensor technology has enabled a novel and sophisticated analytical approach for in situ and simultaneous detection of multiple analytes. Multiplexing offers the possibility of monitoring complex microenvironments, which are otherwise difficult to characterize in a rapid and minimally invasive manner. For example, this technology could be utilized for simultaneous monitoring of extracellular levels of, glucose, lactate and pH,9 which are important metabolic indicators of disease states7,10-14 (e.g., cancer proliferation) and exercise-induced acidosis.15

This article details the construction of a multiplexed microneedle-based sensor. The device is being developed for in situ sampling and electrochemical analysis of multiple analytes in a rapid and selective manner. We envision clinical medicine and biomedical research uses for these microneedle-based sensors.

复透皮电化学传感的空心微针为基础的传感器

The development of a minimally invasive multiplexed monitoring system for rapid analysis of biologically-relevant molecules could offer individuals ...

科研

生物工程

Bacterial Detection &amp; Identification Using Electrochemical Sensors

Electrochemical sensors are widely used for rapid and accurate measurement of blood glucose and can be adapted for detection of a wide variety of analytes. Electrochemical sensors operate by transducing a biological recognition event into a useful electrical signal. Signal transduction occurs by coupling the activity of a redox enzyme to an amperometric electrode. Sensor specificity is either an inherent characteristic of the enzyme, glucose oxidase in the case of a glucose sensor, or a product of linkage between the enzyme and an antibody or probe.
Here, we describe an electrochemical sensor assay method to directly detect and identify bacteria. In every case, the probes described here are DNA oligonucleotides. This method is based on sandwich hybridization of capture and detector probes with target ribosomal RNA (rRNA). The capture probe is anchored to the sensor surface, while the detector probe is linked to horseradish peroxidase (HRP). When a substrate such as 3,3',5,5'-tetramethylbenzidine (TMB) is added to an electrode with capture-target-detector complexes bound to its surface, the substrate is oxidized by HRP and reduced by the working electrode. This redox cycle results in shuttling of electrons by the substrate from the electrode to HRP, producing current flow in the electrode.

We describe an electrochemical sensor assay method for rapid bacterial detection and identification. The assay involves a sensor array functionalized with DNA oligonucleotide capture probes for ribosomal RNA (rRNA) species-specific sequences. Sandwich hybridization of target rRNA with the capture probe and a horseradish peroxidase-linked DNA oligonucleotide detector probe produces a measurable amperometric current.

使用电化学传感器的细菌检测和鉴定

Electrochemical  sensors are widely used for rapid and accurate measurement of blood glucose and  can be adapted for detection of a wide variety of ...

Electric Cell-substrate Impedance Sensing for the Quantification of Endothelial Proliferation, Barrier Function, and Motility

Electric Cell-substrate Impedance Sensing (ECIS) is an in vitro impedance measuring system to quantify the behavior of cells within adherent cell layers. To this end, cells are grown in special culture chambers on top of opposing, circular gold electrodes. A constant small alternating current is applied between the electrodes and the potential across is measured. The insulating properties of the cell membrane create a resistance towards the electrical current flow resulting in an increased electrical potential between the electrodes. Measuring cellular impedance in this manner allows the automated study of cell attachment, growth, morphology, function, and motility. Although the ECIS measurement itself is straightforward and easy to learn, the underlying theory is complex and selection of the right settings and correct analysis and interpretation of the data is not self-evident. Yet, a clear protocol describing the individual steps from the experimental design to preparation, realization, and analysis of the experiment is not available. In this article the basic measurement principle as well as possible applications, experimental considerations, advantages and limitations of the ECIS system are discussed. A guide is provided for the study of cell attachment, spreading and proliferation; quantification of cell behavior in a confluent layer, with regard to barrier function, cell motility, quality of cell-cell and cell-substrate adhesions; and quantification of wound healing and cellular responses to vasoactive stimuli. Representative results are discussed based on human microvascular (MVEC) and human umbilical vein endothelial cells (HUVEC), but are applicable to all adherent growing cells.

This protocol reviews Electric Cell-substrate Impedance Sensing, a method to record and analyze the impedance spectrum of adherent cells for the quantification of cell attachment, proliferation, motility, and cellular responses to pharmacological and toxic stimuli. Detection of endothelial permeability and assessment of cell-cell and cell-substrate contacts are emphasized.

电动细胞基质阻抗传感的内皮细胞增殖，屏障功能的定量和能动性

Electric Cell-substrate Impedance Sensing (ECIS) is an in vitro impedance measuring system to quantify the behavior of cells within adherent cell layers. ...

Preparation of Silicon Nanowire Field-effect Transistor for Chemical and Biosensing Applications

Surveillance using biomarkers is critical for the early detection, rapid intervention, and reduction in the incidence of diseases. In this study, we describe the preparation of polycrystalline silicon nanowire field-effect transistors (pSNWFETs) that serve as biosensing devices for biomarker detection. A protocol for chemical and biomolecular sensing by using pSNWFETs is presented. The pSNWFET device was demonstrated to be a promising transducer for real-time, label-free, and ultra-high-sensitivity biosensing applications. The source/drain channel conductivity of a pSNWFET is sensitive to changes in the environment around its silicon nanowire (SNW) surface. Thus, by immobilizing probes on the SNW surface, the pSNWFET can be used to detect various biotargets ranging from small molecules (dopamine) to macromolecules (DNA and proteins). Immobilizing a bioprobe on the SNW surface, which is a multistep procedure, is vital for determining the specificity of the biosensor. It is essential that every step of the immobilization procedure is correctly performed. We verified surface modifications by directly observing the shift in the electric properties of the pSNWFET following each modification step. Additionally, X-ray photoelectron spectroscopy was used to examine the surface composition following each modification. Finally, we demonstrated DNA sensing on the pSNWFET. This protocol provides step-by-step procedures for verifying bioprobe immobilization and subsequent DNA biosensing application.

We describe key steps for biosensing by using polysilicon nanowire field-effect transistors, including the preparation of the device and the immobilization and confirmation of a DNA molecular probe on the nanowire surface, as well as conditions for DNA sensing.

硅纳米线场效应晶体管的研制化学生物传感与应用

Surveillance using biomarkers is critical for the early detection, rapid intervention, and reduction in the incidence of diseases. In this study, we ...

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and trade one defect for another. Tissue engineering holds the promise to address this increasing demand. However, most tissue engineering strategies fail to generate stable and functional tissue substitutes because of poor vascularization. This paper focuses on an in vivo tissue engineering chamber model of intrinsic vascularization where a perfused artery and a vein either as an arteriovenous loop or a flow-through pedicle configuration is directed inside a protected hollow chamber. In this chamber-based system angiogenic sprouting occurs from the arteriovenous vessels and this system attracts ischemic and inflammatory driven endogenous cell migration which gradually fills the chamber space with fibro-vascular tissue. Exogenous cell/matrix implantation at the time of chamber construction enhances cell survival and determines specificity of the engineered tissues which develop. Our studies have shown that this chamber model can successfully generate different tissues such as fat, cardiac muscle, liver and others. However, modifications and refinements are required to ensure target tissue formation is consistent and reproducible. This article describes a standardized protocol for the fabrication of two different vascularized tissue engineering chamber models in vivo.

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

This is a guideline for constructing in vivo vascularized tissue using a microsurgical arteriovenous loop or a flow-through pedicle configuration inside a tissue engineering chamber. The vascularized tissues generated can be employed for organ regeneration and replacement of tissue defects, as well as for drug testing and disease modeling.

在组织工程由内在血管化<em&gt;在体内</em&gt;组织工程商会

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and ...

Wet Chemistry and Peptide Immobilization on Polytetrafluoroethylene for Improved Cell-adhesion

Endowing materials surface with cell-adhesive properties is a common strategy in biomaterial research and tissue engineering. This is particularly interesting for already approved polymers that have a long standing use in medicine because these materials are well characterized and legal issues associated with the introduction of newly synthesized polymers may be avoided. Polytetrafluoroethylene (PTFE) is one of the most frequently employed materials for the manufacturing of vascular grafts but the polymer lacks cell adhesion promoting features. Endothelialization, i.e., complete coverage of the grafts inner surface with a confluent layer of endothelial cells is regarded key to optimal performance, mainly by reducing thrombogenicity of the artificial interface.
This study investigates the growth of endothelial cells on peptide-modified PTFE and compares these results to those obtained on unmodified substrate. Coupling with the endothelial cell adhesive peptide Arg-Glu-Asp-Val (REDV) is performed via activation of the fluorin-containing polymer using the reagent sodium naphthalenide, followed by subsequent conjugation steps. Cell culture is accomplished using Human Umbilical Vein Endothelial Cells (HUVECs) and excellent cellular growth on peptide-immobilized material is demonstrated over a two-week period.

Cell-adhesiveness is key to many approaches in biomaterial research and tissue engineering. A step-by-step technique is presented using wet-chemistry for the surface modification of the important polymer PTFE with peptides.

湿化学和肽固定在聚四氟乙烯为改善细胞粘附

Endowing materials surface with cell-adhesive properties is a common strategy in biomaterial research and tissue engineering. This is particularly ...

Development of an Electrochemical DNA Biosensor to Detect a Foodborne Pathogen

Vibrio parahaemolyticus (V. parahaemolyticus) is a common foodborne pathogen that contributes to a large proportion of public health problems globally, significantly affecting the rate of human mortality and morbidity. Conventional methods for the detection of V. parahaemolyticus such as culture-based methods, immunological assays, and molecular-based methods require complicated sample handling and are time-consuming, tedious, and costly. Recently, biosensors have proven to be a promising and comprehensive detection method with the advantages of fast detection, cost-effectiveness, and practicality. This research focuses on developing a rapid method of detecting V. parahaemolyticus with high selectivity and sensitivity using the principles of DNA hybridization. In the work, characterization of synthesized polylactic acid-stabilized gold nanoparticles (PLA-AuNPs) was achieved using X-ray Diffraction (XRD), Ultraviolet-visible Spectroscopy (UV-Vis), Transmission Electron Microscopy (TEM), Field-emission Scanning Electron Microscopy (FESEM), and Cyclic Voltammetry (CV). We also carried out further testing of stability, sensitivity, and reproducibility of the PLA-AuNPs. We found that the PLA-AuNPs formed a sound structure of stabilized nanoparticles in aqueous solution. We also observed that the sensitivity improved as a result of the smaller charge transfer resistance (Rct) value and an increase of active surface area (0.41 cm2). The development of our DNA biosensor was based on modification of a screen-printed carbon electrode (SPCE) with PLA-AuNPs and using methylene blue (MB) as the redox indicator. We assessed the immobilization and hybridization events by differential pulse voltammetry (DPV). We found that complementary, non-complementary, and mismatched oligonucleotides were specifically distinguished by the fabricated biosensor. It also showed reliably sensitive detection in cross-reactivity studies against various food-borne pathogens and in the identification of V. parahaemolyticus in fresh cockles.

A protocol for the development of an electrochemical DNA biosensor comprising a polylactic acid-stabilized, gold nanoparticles-modified, screen-printed carbon electrode to detect Vibrio parahaemolyticus is presented.

电化学 DNA 生物传感器检测食源性致病菌的研究进展

Vibrio parahaemolyticus (V. parahaemolyticus) is a common foodborne pathogen that contributes to a large proportion of public health problems globally, ...

In Vitro Multiparametric Cellular Analysis by Micro Organic Charge-modulated Field-effect Transistor Arrays

Modern electrophysiology has been constantly fueled by the parallel development of increasingly sophisticated tools and materials. In turn, discoveries in this field have driven technological progress in a back-and-forth process that ultimately determined the impressive achievements of the past 50 years. However, the most employed devices used for cellular interfacing (namely, the microelectrode arrays and microelectronic devices based on transistors) still present several limitations such as high cost, the rigidity of the materials, and the presence of an external reference electrode. To partially overcome these issues, there have been developments in a new scientific field called organic bioelectronics, resulting in advantages such as lower cost, more convenient materials, and innovative fabrication techniques.
Several interesting new organic devices have been proposed during the past decade to conveniently interface with cell cultures.&#160;This paper presents the protocol for the fabrication of devices for cellular interfacing based on the organic charge-modulated field-effect transistor (OCMFET). These devices, called micro OCMFET arrays (MOAs), combine the advantages of organic electronics and the peculiar features of the OCMFET to prepare transparent, flexible, and reference-less tools with which it is possible to monitor both the electrical and the metabolic activities of cardiomyocytes and neurons in vitro, thus allowing a multiparametric evaluation of electrogenic cell models.

In Vitro Multiparametric Cellular Analysis by Micro Organic Charge-modulated Field-effect Transistor Arrays

Here, we present the fabrication protocol of an organic charge-modulated field-effect transistor (OCMFET)-based device for in vitro cellular interfacing. The device, called a micro OCMFET array, is a flexible, low-cost, and reference-less device, which will enable the monitoring of the electrical and metabolic activities of electroactive cell cultures.

体外 通过微有机电荷调制场效应晶体管阵列进行多参数蜂窝分析

Modern electrophysiology has been constantly fueled by the parallel development of increasingly sophisticated tools and materials. In turn, discoveries in ...

Disruption of the Blood-Spinal Cord Barrier Using Low-Intensity Focused Ultrasound in a Rat Model

Low-intensity focused ultrasound (LIFU) uses ultrasonic pulsations at lower intensities than ultrasound and is being tested as a reversible and precise neuromodulatory technology. Although LIFU-mediated blood-brain barrier (BBB) opening has been explored in detail, no standardized technique for blood-spinal cord barrier (BSCB) opening has been established to date. Therefore, this protocol presents a method for successful BSCB disruption using LIFU sonication in a rat model, including descriptions of animal preparation, microbubble administration, target selection&#160;and localization, as well as BSCB disruption visualization and confirmation. The approach reported here is particularly useful for researchers who need a fast and cost-effective method to test and confirm target localization and precise BSCB disruption in a small animal model with a focused ultrasound transducer, evaluate the BSCB efficacy of sonication parameters, or explore applications for LIFU at the spinal cord, such as drug delivery, immunomodulation, and neuromodulation. Optimizing this protocol for individual use is recommended, especially for advancing future preclinical, clinical, and translational work.

Disruption of the blood-spinal cord barrier (BSCB) can be successfully achieved with the intravenous administration of microbubbles and the application of low-intensity focused ultrasound (LIFU). This protocol details the opening of the BSCB using LIFU in a rodent model, including equipment setup, microbubble injection, target localization, and BSCB disruption visualization.

在大鼠模型中使用低强度聚焦超声破坏血脊髓屏障

Low-intensity focused ultrasound (LIFU) uses ultrasonic pulsations at lower intensities than ultrasound and is being tested as a reversible and precise ...

通过电电池-衬底传感评估 MOF 诱导的电池行为变化

Electric Cell-Substrate Sensing for Real-Time Evaluation of Metal-Organic Framework Toxicological Profiles

Metal-organic frameworks (MOFs) are hybrids formed through the coordination of metal ions and organic linkers in organic solvents. The implementation of MOFs in biomedical and industrial applications has led to concerns regarding their safety. Herein, the profile of a selected MOF, a zeolitic imidazole framework, was evaluated upon exposure to human lung epithelial cells. The platform for evaluation was a real-time technique (i.e., electric cell-substrate impedance sensing [ECIS]). This study identifies and discusses some of the deleterious effects of the selected MOF on the exposed cells. Furthermore, this study demonstrates the benefits of using the real-time method versus other biochemical assays for comprehensive cell evaluations. The study concludes that observed changes in cell behavior could hint at possible toxicity induced upon exposure&#160;to MOFs of different physicochemical characteristics and the dosage of those frameworks being used. By understanding changes in cell behavior, one foresees the ability to improve safe-by-design strategies of MOFs to be used for biomedical applications by specifically tailoring their physicochemical characteristics.

作者聚焦：评估人肺上皮细胞对金属有机框架反应的进展 

The following study evaluates the toxicological profile of a selected metal-organic framework utilizing electric cell-substrate impedance sensing (ECIS), a real-time, high-throughput screening technique.

用于实时评估金属有机框架毒理学特征的电池-基底传感

Metal-organic frameworks (MOFs) are hybrids formed through the coordination of metal ions and organic linkers in organic solvents. The implementation of ...