The authors had no funding provided for this study.

1.样品架的构建<ol><li>构造由不锈钢隔板和M3的不锈钢牙螺钉，代替与M3的六角螺母保持的样品架。 </li><li>用钳子取出螺纹螺钉的头部和抛光切段维持螺纹图案。 </li><li>当所有单独的组件已经准备好，组装电极夹。每个电极夹包含从而导致11.5厘米手柄的M3螺丝连接在一起的三个间隔。放置六角螺母在螺杆和隔板的接合以锁定连接。 </li><li>焊料（60/40％的Sn / Pb）无牙鳄鱼夹到在杆的端部的螺钉。这将确保牢牢把握分析过程中稍后再连接电极。 </li><li>一旦电极座组装，适用停止过漆（电气密封胶）的多道涂层，以防止不锈钢棒被腐蚀，而沉浸在腐蚀室中。 <ol><li>将所有电极持有人连接到鳄鱼夹入前涂通风柜样本。放置20毫升注射器进入通风柜。用注射器收集停止过漆。 </li><li>打开通风柜，倒停过漆成一个小玻璃瓶。拉10毫升停止断漆到注射器和涂布电极夹头的表面上。确保不覆盖测试样品，这是将要分析的腐蚀。 </li><li>在通风柜每个电极架和地点的涂层一半涂覆，另一半之前干燥。这将有助于获得一个完整的密封良好涂层而不损坏要被涂敷的区域。确保在干燥阶段，新涂的区域不碰其他表面，因为这会毁了应用外衣。 </li><li>放在一个高的位置，电极持有人不接触任何表面的干燥过程。外套由于停止过漆的快速凝固电极快。这样就完成了第一层。 </li><li>一旦干燥，重复该过程，得到3外套沿整个区域。 </li></ol></li><li>在开始运行腐蚀之前，离开人最后一层完成后晾干24小时。所有涂层过程发生在RT，不加热或冷却步骤需要，尽管它们可能加速/减速固化过程。 </li><li> 制造法拉第笼 <ol><li>通过涂布相同大小的两个塑料容器4层铝箔覆盖所有侧面构造的法拉第笼。 </li><li>切两个小孔列于上部塑料容器的边缘，以允许所述电极连接到恒电位仪和氮气管线的氮气罐通过。法拉第笼的分割设计允许在运行结束要删除的上部件，而无需更换下部部分容纳罐。 </li><li>适合外室（水室）插入法拉第笼。 LEAVE在仅当腐蚀容器已经被密封的（在后面的步骤）的下隔室的顶部下半年侧和地点。 </li></ol></li></ol> 2.玻璃器皿清洗<ol><li>每腐蚀运行前清理腐蚀容器（700毫升圆柱瓶）。擦洗家用洗涤剂的容器中，用自来水彻底冲洗。重复此步骤3次。 </li><li>冲洗腐蚀容器用去离子水（DI）水洗涤3次以除去自来水中发现的潜在的污染物。 </li><li>一旦用DI水漂洗完成倾300毫升95％乙醇的进腐蚀容器，摇动周围接触所有内表面。倒出乙醇，重复此步骤3次。 </li><li>发表通风橱下的耐蚀容器30分钟，以允许所有的乙醇完全蒸发。 </li><li>取干净，干燥腐蚀容器中，并与将被用于腐蚀运行电解质冲洗。前面ACH冲洗填充用200ml电解质的腐蚀容器中，并重复此过程3次。在这项研究中，漂洗用磷酸盐缓冲盐水（PBS）的腐蚀容器中。 PBS的通篇使用（10升）的电解质的化学组成为80克NaCl，11.5克Na 2 HPO 4，2克氯化钾和2g KHPO 4。 </li><li>以下漂洗，填充用PBS的所需体积准备用于反应的腐蚀容器中。 </li></ol> 3.设备的安装<ol><li>夹持加热器与一个内置循环系统用夹具的外隔室的一侧。外室的大小需要是大约30厘米×20厘米×20厘米，由玻璃或聚合物的以能够壳体较小腐蚀容器和加热器系统。 </li><li>填用自来水外室，直到水的水平高于腐蚀容器内的悬浮电极的高度高。较小的隔室是腐蚀容器（以前在第2描述的）。 </li><li>用密封的玻璃反应盖腐蚀容器夹，以确保防水密封。腔室的盖子提供了实验和测定装置6的入口点。 </li><li>暂停从反应盖的入口点中的一个温度计，以提供腐蚀电池内部的温度的读数。暂停从盖子使用其他3个入口点的所有三个电极。使用聚四氟乙烯（PTFE）胶带，以确保每个连接的密封。 </li><li>使用三电极结构，包括一个参考，计数器，和工作电极。工作电极是不锈钢螺钉（下分析样品）。插入电极插入腐蚀容器之前，擦拭用80％的乙醇浸泡在填充用100ml的PBS的玻璃烧杯擦拭和地点。 </li><li>使用连接引脚到电极夹附着到电极上吊带。适合该电极Ë吊带进入腐蚀容器的盖子的入口点。 </li><li>放置在工作电极中央与计数器和参考电极从任一侧被暂停。密封玻璃入口，并使用聚四氟乙烯带腐蚀吊带。 </li><li>为参考电极，使用一个标准的Ag / AgCl。对电极，使用铂网状这是松散弯曲包裹试样围绕被测（工作电极）。 </li><li>填用吸管用3M氯化钾的银/氯化银电极。经过广泛的使用，变更和填充银银/氯。要做到这一点释放电极尖端空出的流体成一个小玻璃收集容器（烧杯）。一旦所有的溶液是去除插入尖端和用3M氯化钾填充。 </li><li>使用上的所有路口磁带确保整个室是密封的。 </li><li>一旦该腔室被放置在腐蚀容器内所有电极密封，将温度设定为37℃，并打开氮气阀机智哈流量为150 立方厘米 / min的速率。在进行运行前留下60分钟的温度和氮气运转。保持氮气运行在实验的持续时间。 </li></ol> 4.运行腐蚀试验<ol><li>打开电化学软件包，其与USB控制电位接口。 </li><li>制作和3个电极恒电位仪之间的电气连接，然后打开电位上。 </li><li>打开并使用"测量视图"来查看腐蚀环境的潜力和电流读数。期间在没有斜坡电位尚未施加工作（正电位）和计数器之间的电流读数的开路电势（OCP）相（负）电极是围绕（0±0.01）微安。用PTFE带腔室的不当密封可能会导致在当前读数的波动，由于该腔室被用氮气充气至除去氧分子。 </li><li>离开样以平衡，腐蚀容器环境中稳定。这个持续时间而变化（1至6小时），并依赖于材料。监视用的测量，以确定是否达到了稳定的条件的可能性。一旦达到稳定的条件下的电位将与无波动恒定。 </li><li>后稳定条件达到，开始腐蚀运行。不过在此之前可以做到的，填写"腐蚀计划"和"循环伏安法（CV）"的条件下利用分析软件提供的框架模板。 </li><li>从程序选项卡中设置视图中的循环伏安恒电位的过程。 </li><li>启用以下参数腐蚀运行进行采样:时间，工作电极（WE）电位，电流腐蚀运行。 </li><li>选择自动电流范围的选项。设置的最高电流的范围为10毫安，并在R的最低电流安格成为了WE 10 nA的。 </li><li>确保最终截止选择由设置&#39;循环回&#39;参数至0.8毫伏，以允许磁滞回线来完成通过电位控制。 </li><li>从测量视图进入OCP参数文本框中记录OCP。设置下面的记录OCP值开始潜力100毫伏。设置上限顶点电位至800毫伏，下部顶点到100毫伏下开始电位和停止电势为100毫伏低于下顶点潜力。设置扫描速率到0.001伏/秒和步潜力0.0024 V /秒。现在，按启动键。 </li></ol> 5.腐蚀运行完成后注意:在腐蚀结束运行偏振扫描软件的分析视图中示出。对于每一个极化运行演示者视图列出了OCP，该地块对于E与T和CV楼梯，为E主场迎战日志（I）的情节。 <ol><li>在每个情节升墨，确定数据点的内部过滤，塔菲尔外推，和情节的选项。展开向显示感兴趣的各种参数，它们共同形成了电化学参数的每一个环节。极化扫描（电流密度-电势），确定开路电位，点蚀电位（E 坑 ）和保护电势（E 亲 ）。 </li><li>制表使用塔菲尔斜率链路的腐蚀速率下的阳极和阴极塔菲尔常数，腐蚀速率，腐蚀电流，腐蚀电流密度，起始电位，结束电位。 </li></ol> 6.删除从电极保持样品<ol><li>准备在通风橱下50毫升二氯甲烷3小罐。 </li><li>由浸渍保持件的下端的二氯甲烷为一个通风橱内30分钟除去由电极支架测试的样品。 </li><li>一旦分离，将试样进入下一个罐子二氯甲烷和离开15分钟。重复此过程，与第三和最后冲洗摆脱在样品上的安装部分的任何多余的涂料。 </li><li>擦拭从剪辑和样本剩余的密封胶，最后用去离子水冲洗。 </li></ol>

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M. a. t. e. r. .. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+corrosion+of+modular+hip+prosthesis+components+in+mixed+and+similar+metal+combinations.+The+effect+of+crevice,+stress,+motion+and+allot+coupling.">In vivo corrosion of modular hip prosthesis components in mixed and similar metal combinations. The effect of crevice, stress, motion and allot coupling.</a> J. Biomed. Mater. Res. 76 (1), 1533-1544 (1993).</li><li>Browne, J. A., Bechtold, C. D., Berry, D. J., Hanssen, A. D., Lewallen, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Failed+metal-on-metal+hip+arthroplasties:+a+spectrum+of+clinical+presentations+and+operative+findings.">Failed metal-on-metal hip arthroplasties: a spectrum of clinical presentations and operative findings.</a> Clin. Orthop. Relat. Res. 468 (9), 2313-2320 (2010).</li><li>Jantzen, C., Jorgensen, H. L., Duus, B. R., Sporring, S. L., Lauritzen, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chromium+and+cobalt+ion+concentrations+in+blood+and+serum+following+various+types+of+metal-on-metal+hip+arthroplasties:+a+literature+overview.">Chromium and cobalt ion concentrations in blood and serum following various types of metal-on-metal hip arthroplasties: a literature overview.</a> Acta Orthop. 84 (3), 229-236 (2013).</li><li>Frangini, S., De Cristofaro, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+galvanostatic+polarisation+method+for+determining+reliable+pitting+potentials+on+stainless+steels+in+crevice-free+conditions.">Analysis of galvanostatic polarisation method for determining reliable pitting potentials on stainless steels in crevice-free conditions.</a> Corros Sci. 45 (12), 2769-2786 (2002).</li></ol>

The authors have nothing to disclose.

从不锈钢试样产生偏振扫描显示清洁连续曲线，在指示运作良好的腐蚀系统，该系统既可靠和可再现29的文献看出扫描相关联。差电位点蚀电位的再现性是确定与几百毫伏的蔓延，与孔蚀电位的特征在于一个随机过程29。这通常是由于温度，卤化物含量和电势（V）的变量;因此，从实际安装在E 科尔获得较小的变化是指示协议，并具有改善体外设置上述调整。 在过程中的一个关键步骤是建立在反应器内稳定的环境，降低噪音。创建和之前每次运行以下具体步骤来清洁反应容器中改进的结果和公关ovided重现性好，可靠的读数。电解质内的污染物可以改变腐蚀环境和材料的腐蚀的反应，引起在结果不符。最小化此被发现是在协议的关键步骤。到位的清洗程序，在电极和腐蚀容器消除潜在的杂质，这本来是以前见过的差异的一个因素。 在过程中的第二个关键步骤是提供一个电屏蔽的样品架，以消除该腔室中的任何金属的接触。从任何电化学传导性完全屏蔽金属持有人的意义是为了防止外部金属的干扰。在不脱离金属物种的任何其它形式的待测分离该金属试样的腐蚀分析不能提供试样的准确读数。如果持有人不能正确地涂，他们会腐蚀。如果科尔osion看出在不属于被检查的读数不能用于分析和将需要另一次操作的金属部件。 最初在现场录音相位扫描显示出振荡，这可能是由于不稳定或高噪声电平。在运行过程中的振动被视为不稳定的结果。这是由于该电位的失败，以保持细胞的潜在30的控制。由于高噪声电平的振荡可以从外部源，这需要一定程度的过滤。故障排除的关键是连接电解质和电极之间的陶瓷电容盘。抑制电容器通常掺入到交变电流线路滤波器来抑制这通常通过电气/电子设备产生的电磁或无线电频率的干扰，以及减少电气开关噪声。陶瓷CAPA四个不同大小citors被用来分析极化曲线上其噪声抑制功效，为0.001至1μF。 0.1μF电容显著平滑极化曲线。所有的噪音被抑制;除去原始扫描中找到的所有峰值。实验结果表明，电感开始降低1μF电容器的噪声抑制功效，同时不影响在本噪声的频率范围的0.1 F。 动电位腐蚀将为材料控制环境提供了一个体外腐蚀试验系统。材质的防腐能力，可按照该材料制成的任何形式的操纵进行评估。腐蚀与能力的分析来控制不同的参数将提供进一步的检查和对金属材料的腐蚀的变化进行分析。该协议同时具有局限性和优势。这样做的意义满足HOD相对于其它的方法是相对较低的成本和快速的过程来执行复杂的分析1,4,5。该协议将提供实验室检测的可靠来源进行。然而该协议的一个限制是，可以在一个点上被测试平行样品的数量有限。设置仅提供每个测试一个样品，这会延长试验时间为大量的样品。

电化学技术提供了快速且相对便宜的方法获得的材料的电化学性能。这些技术在通过观察电荷转移过程的受控电化学干扰1-5的响应，以检测金属的腐蚀的能力，主要是基础的。体的环境中的金属植入物的腐蚀是非常关键的，由于在生物相容性和材料完整性6的不利影响。促进身体内植入物的腐蚀的主要因素是表面氧化，导致金属离子7-11的释放增多的溶解。这将导致不良的生物反应，其可以发现局部的，但具有潜在的全身效应，导致植入物10,12-28的过早失效。 试样的腐蚀特性从产生的偏振扫描预测由恒电位。偏振扫描允许对金属基板的动能和腐蚀参数的推断。在扫描期间，氧化或还原的电活性物种可以通过电荷转移和反应物或产物的运动受到限制。这些因素都是由偏振扫描包封;因此具有在多个周期产生一个可靠和可重复的偏振扫描系统的重要性是非常重要的。这份手稿的重点是提供一个确定的协议，以获得一个运作良好的电位腐蚀体系所采取的基本原理和步骤。

<table border="0" cellpadding="0" cellspacing="0" width="573">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Potentiostat</td>
			<td style="width:71px;">Metrohm</td>
			<td style="width:119px;">PGSTAT101</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Ag/AgCl reference electrode, shielded</td>
			<td style="width:71px;">Metrohm</td>
			<td style="width:119px;">6.0729.100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Electrode shaft</td>
			<td style="width:71px;">Metrohm</td>
			<td style="width:119px;">6.1241.060</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Polisher Forcipol 1v</td>
			<td style="width:71px;">Metkon</td>
			<td align="right" style="width:119px;">3602</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Clindrical flask 700mL</td>
			<td style="width:71px;">SciLabware</td>
			<td style="width:119px;">FR700F</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Reaction lid</td>
			<td style="width:71px;">SciLabware</td>
			<td style="width:119px;">MAF2/41</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Dichloromethane</td>
			<td style="width:71px;">Sigma-Aldrich</td>
			<td style="width:119px;">MKBR7629V</td>
			<td>use under a fumehood. Wear protective clothing</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Thermo / HAAKE D Series Immersion Circulators</td>
			<td style="width:71px;">Haake</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

potentiodynamic corrosion testing

Different metallic materials have different polarization characteristics as dictated by the open circuit potential, breakdown potential, and passivation potential of the material. The detection of these electrochemical parameters identifies the corrosion factors of a material. A reliable and well-functioning corrosion system is required to achieve this.
Corrosion of the samples was achieved via a potentiodynamic polarization technique employing a three-electrode configuration, consisting of reference, counter, and working electrodes. Prior to commencement a baseline potential is obtained. Following the stabilization of the corrosion potential (Ecorr), the applied potential is ramped at a slow rate in the positive direction relative to the reference electrode. The working electrode was a stainless steel screw. The reference electrode was a standard Ag/AgCl. The counter electrode used was a platinum mesh. Having a reliable and well-functioning in vitro corrosion system to test biomaterials provides an in-expensive technique that allows for the systematic characterization of the material by determining the breakdown potential, to further understand the material's response to corrosion. The goal of the protocol is to set up and run an in vitro potentiodynamic corrosion system to analyze pitting corrosion for small metallic medical devices.

在该过程结束体外腐蚀系统设置进行腐蚀研究。如腐蚀容器和法拉第笼的清洁特定程序被引入协议来改善噪声性能。一个良好的偏振扫描的基本概念是，以确定以理解的金属材料的腐蚀敏感性提供有价值的信息，该材料的电物理条件。的程序和协议，以实现可靠的和可重复的结果是至关重要的。获得其上的优化可以帮助，可以在使用过程中出现的某些问题的信息，识别并添加到当前的程序。预先存在的非校准系统产生的偏振扫描（ 图1）。该扫描显示了散射点的集合，其不直接允许电物理COND的测定被检查物质的itions。在OCP的识别是不切合实际和腐蚀或再钝化电位难以阅读。与预先存在的设置中发现的故障包括偏振扫描没有达到的最大潜力，并过早地切断由于高噪声水平。其次在现场录音相位扫描显示出振荡，这可能是由于不稳定或高噪声水平。在运行过程中的振荡被视为不稳定的结果。最后连续扫描不会交付重复性的结果使得不可能确定特定材料的电物理性能。 <img alt="图1" src="/files/ftp_upload/54351/54351fig1.jpg" /> 图1. 接受电动电位极化后Nitanol样品的极化扫描。该图表示一个嘈杂情节第在不提供腐蚀参数的准确诠释。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54351/54351fig1large.jpg" target="_blank">请点击此处查看这个数字的原始版本。</a> 在噪声性能的提高示于（ 图2）。扫描显示了向 ​​前的方向和反向扫描并指定在所述的保护电势（E 亲 ）和点蚀电位（E 坑 ）观察到的点。情节是干净，在整个扫从而可以观察到清晰的整个范围内无噪音或差异。的滞后在设定的电位反转，返回到拦截阳极曲线，确定保护电位。极化扫描和极化曲线是输出，提供所需的基本参数。这些参数从偏振扫描确定的，因此，甲肝ING提供高质量的扫描哪些是可再生的，可靠的识别可以推断参数前重要的系统。 <img alt="图2" src="/files/ftp_upload/54351/54351fig2.jpg" /> 图2. 不锈钢316偏振扫描这是一个明确的情节类似偏振扫描以下腐蚀其中感兴趣的参数可以很容易地检测出来。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54351/54351fig2large.jpg" target="_blank">请点击此处查看这个数字的原始版本。</a> 进行研究分析以下点蚀在金属螺丝表面形貌变化。从研究中获得的均值为E 科尔值为（-0.414±0.05）V.为样本的平均点蚀电位为（0.49±0.12）V，这是智慧欣极化曲线的活性区域。样品的平均保护电位为（-0.16±0.02）V.每个螺丝形成沿着所述表面在确认从出详细的地形变化宏观图像的结果，由于凹坑的形成和凹坑内的变化的局部凹坑（ 图3）。该材料的表面形貌的定量表明，该材料的粗糙度已经降低作为整体的表面粗糙度;螺丝中 ，R a为（159.9±7.3）微米（无腐蚀）和（124.7±18.3）微米（腐蚀）。为腐蚀试样相比，非腐蚀的R A是显著降低（P = 0.02）。平均最大高度- [R Z可以（469.3±16.5）微米（不熔蚀）和（683.2±85.8）微米（腐蚀）标识要腐蚀和非腐蚀样品之间的高度变化的显著差异（p = 0.04）。平均最大HEIGHT（R T）降低在（502.61±51.2）微米相比，腐蚀螺钉（592.23±119.7）微米无腐蚀。 <img alt="图3" src="/files/ftp_upload/54351/54351fig3.jpg" /> 图3. 从该展示的局部上的螺纹表面腐蚀的光学轮廓图像。腐蚀已都在表面的谷和波谷被检测到。点蚀的目测成功腐蚀设置来实现的。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54351/54351fig3large.jpg" target="_blank">请点击此处查看这个数字的原始版本。</a>

Watch this Scientific Journal Video about Potentiodynamic Corrosion Testing at JoVE.com

Potentiodynamic Corrosion Testing

Here, we present a protocol to set up and run an in vitro potentiodynamic corrosion system to analyze pitting corrosion for small metallic medical devices.

动电位腐蚀试验

Different metallic materials have different polarization characteristics as dictated by the open circuit potential, breakdown potential, and passivation ...

Nanyang Technological University

Research

JoVE Journal

Environment

17.5K 次观看.  Prince of Wales Clinical School. Here, we present a protocol to set up and run an in vitro potentiodynamic corrosion system to analyze pitting corrosion for small metallic medical devices. 

视频: Potentiodynamic Corrosion Testing

Preparation and Testing of Plant Seed Meal-based Wood Adhesives

Recently, the interest in plant seed meal-based products as wood adhesives has steadily increased, as these plant raw materials are considered renewable and environment-friendly. These natural products may serve as alternatives to petroleum-based adhesives to ease environmental and sustainability concerns. This work demonstrates the preparation and testing of the plant seed-based wood adhesives using cottonseed and soy meal as raw materials. In addition to untreated meals, water washed meals and protein isolates are prepared and tested. Adhesive slurries are prepared by mixing a freeze-dried meal product with deionized water (3:25 w/w) for 2 hr. Each adhesive preparation is applied to one end of 2 wood veneer strips using a brush. The tacky adhesive coated areas of the wood veneer strips are lapped and glued by hot-pressing. Adhesive strength is reported as the shear strength of the bonded wood specimen at break. Water resistance of the adhesives is measured by the change in shear strength of the bonded wood specimens at break after water soaking. This protocol allows one to assess plant seed-based agricultural products as suitable candidates for substitution of synthetic-based wood adhesives. Adjustments to the adhesive formulation with or without additives and bonding conditions could optimize their adhesive properties for various practical applications.

To facilitate the effort in seeking more economic and environment-friendly formulations of natural product-based wood adhesives, this work demonstrates the preparation and testing of plant seed-based wood adhesives. This protocol allows one to assess plant seed-based agricultural products as suitable candidates for the substitution of synthetic-based wood adhesives.

准备和植物种子膳食为基础的木材胶粘剂的测试

Recently, the interest in plant seed meal-based products as wood adhesives has steadily increased, as these plant raw materials are considered renewable ...

科研

环境科学

Protocols for Robust Herbicide Resistance Testing in Different Weed Species

Robust protocols to test putative herbicide resistant weed populations at whole plant level are essential to confirm the resistance status. The presented protocols, based on whole-plant bioassays performed in a greenhouse, can be readily adapted to a wide range of weed species and herbicides through appropriate variants. Seed samples from plants that survived a field herbicide treatment are collected and stored dry at low temperature until used. Germination methods differ according to weed species and seed dormancy type. Seedlings at similar growth stage are transplanted and maintained in the greenhouse under appropriate conditions until plants have reached the right growth stage for herbicide treatment. Accuracy is required to prepare the herbicide solution to avoid unverifiable mistakes. Other critical steps such as the application volume and spray speed are also evaluated. The advantages of this protocol, compared to others based on whole plant bioassays using one herbicide dose, are related to the higher reliability and the possibility of inferring the resistance level. Quicker and less expensive in vivo or in vitro diagnostic screening tests have been proposed (Petri dish bioassays, spectrophotometric tests), but they provide only qualitative information and their widespread use is hindered by the laborious set-up that some species may require. For routine resistance testing, the proposed whole plant bioassay can be applied at only one herbicide dose, so reducing the costs.

A robust and flexible approach to confirm herbicide resistance in weed populations is presented. This protocol allows the herbicide resistance levels to be inferred and applied to a wide range of weed species and herbicides with minor adaptations.

协议的鲁棒抗除草剂试验在不同杂草

Robust protocols to test putative herbicide resistant weed populations at whole plant level are essential to confirm the resistance status. The presented ...

A CO2 Concentration Gradient Facility for Testing CO2 Enrichment and Soil Effects on Grassland Ecosystem Function

Continuing increases in atmospheric carbon dioxide concentrations (CA) mandate techniques for examining impacts on terrestrial ecosystems. Most experiments examine only two or a few levels of CA concentration and a single soil type, but if CA can be varied as a gradient from subambient to superambient concentrations on multiple soils, we can discern whether past ecosystem responses may continue linearly in the future and whether responses may vary across the landscape. The Lysimeter Carbon Dioxide Gradient Facility applies a 250 to 500 &#181;l L-1 CA gradient to Blackland prairie plant communities established on lysimeters containing clay, silty clay, and sandy soils. The gradient is created as photosynthesis by vegetation enclosed in in temperature-controlled chambers progressively depletes carbon dioxide from air flowing directionally through the chambers. Maintaining proper air flow rate, adequate photosynthetic capacity, and temperature control are critical to overcome the main limitations of the system, which are declining photosynthetic rates and increased water stress during summer. The facility is an economical alternative to other techniques of CA enrichment, successfully discerns the shape of ecosystem responses to subambient to superambient CA enrichment, and can be adapted to test for interactions of carbon dioxide with other greenhouse gases such as methane or ozone.

A CO2 Concentration Gradient Facility for Testing CO2 Enrichment and Soil Effects on Grassland Ecosystem Function

The Lysimeter Carbon Dioxide Gradient Facility creates a 250 to 500 &#181;l L-1 linear carbon dioxide gradient in temperature-controlled chambers housing grassland plant communities on clay, silty clay, and sandy soil monoliths. The facility is used to determine how past and future carbon dioxide levels affect grassland carbon cycling.

一个外线<sub&gt; 2</sub&gt;浓度梯度用于测试CO<sub&gt; 2</sub&gt;浓度与土壤对草地生态系统功能的影响

Continuing increases in atmospheric carbon dioxide concentrations (CA) mandate techniques for examining impacts on terrestrial ecosystems. Most ...

Preparation and Testing of Impedance-based Fluidic Biochips with RTgill-W1 Cells for Rapid Evaluation of Drinking Water Samples for Toxicity

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity sensor. A monolayer of RTgill-W1 cells forms on the sensing electrodes enclosed within the biochips. The biochips are then used for testing in a field portable electric cell-substrate impedance sensing (ECIS) device designed for rapid toxicity testing of drinking water. The manuscript further describes how to run a toxicity test using the prepared biochips. A control water sample and the test water sample are mixed with pre-measured powdered media and injected into separate channels of the biochip. Impedance readings from the sensing electrodes in each of the biochip channels are measured and compared by an automated statistical software program. The screen on the ECIS instrument will indicate either "Contamination Detected" or "No Contamination Detected" within an hour of sample injection. Advantages are ease of use and rapid response to a broad spectrum of inorganic and organic chemicals at concentrations that are relevant to human health concerns, as well as the long-term stability of stored biochips in a ready state for testing. Limitations are the requirement for cold storage of the biochips and limited sensitivity to cholinesterase-inhibiting pesticides. Applications for this toxicity detector are for rapid field-portable testing of drinking water supplies by Army Preventative Medicine personnel or for use at municipal water treatment facilities.

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial cells for use in a field portable electric cell-substrate impedance sensor. The protocol for running a rapid drinking water toxicity test with the sensor is also described.

准备和测试阻抗为基础与RTgill-W1细胞饮用水进行快速评估水样进行毒性射流生物芯片

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity ...

Dispersion of Nanomaterials in Aqueous Media: Towards Protocol Optimization

The sonication process is commonly used for de-agglomerating and dispersing nanomaterials in aqueous based media, necessary to improve homogeneity and stability of the suspension. In this study, a systematic step-wise approach is carried out to identify optimal sonication conditions in order to achieve a stable dispersion. This approach has been adopted and shown to be suitable for several nanomaterials (cerium oxide, zinc oxide, and carbon nanotubes) dispersed in deionized (DI) water. However, with any change in either the nanomaterial type or dispersing medium, there needs to be optimization of the basic protocol by adjusting various factors such as sonication time, power, and sonicator type as well as temperature rise during the process. The approach records the dispersion process in detail. This is necessary to identify the time points as well as other above-mentioned conditions during the sonication process in which there may be undesirable changes, such as damage to the particle surface thus affecting surface properties. Our goal is to offer a harmonized approach that can control the quality of the final, produced dispersion. Such a guideline is instrumental in ensuring dispersion quality repeatability in the nanoscience community, particularly in the field of nanotoxicology.

Here, we present a step-wise protocol for the dispersion of nanomaterials in aqueous media with real-time characterization to identify the optimal sonication conditions, intensity, and duration for improved stability and uniformity of nanoparticle dispersions without impacting the sample integrity.

纳米材料在水介质中的分散: 对协议优化的研究

The sonication process is commonly used for de-agglomerating and dispersing nanomaterials in aqueous based media, necessary to improve homogeneity and ...

On the Preparation and Testing of Fuel Cell Catalysts Using the Thin Film Rotating Disk Electrode Method

We present a step-by-step tutorial to prepare proton exchange membrane fuel cell (PEMFC) catalysts, consisting of Pt nanoparticles (NPs) supported on a high surface area carbon, and to test their performance in thin film rotating disk electrode (TF-RDE) measurements. The TF-RDE methodology is widely used for catalyst screening; nevertheless, the measured performance sometimes considerably differs among research groups. These uncertainties impede the advancement of new catalyst materials and, consequently, several authors discussed possible best practice methods and the importance of benchmarking.
The visual tutorial highlights possible pitfalls in the TF-RDE testing of Pt/C catalysts. A synthesis and testing protocol to assess standard Pt/C catalysts is introduced that can be used together with polycrystalline Pt disks as benchmark catalysts. In particular, this study highlights how the properties of the catalyst film on the glassy carbon (GC) electrode influence the measured performance in TF-RDE testing. To obtain thin, homogeneous catalyst films, not only the catalyst preparation, but also the ink deposition and drying procedures are essential. It is demonstrated that an adjustment of the ink&#39;s pH might be necessary, and how simple control measurements can be used to check film quality. Once reproducible TF-RDE measurements are obtained, determining the Pt loading on the catalyst support (expressed as Pt wt%) and the electrochemical surface area is necessary to normalize the determined reaction rates to either surface area or Pt mass. For the surface area determination, so-called CO stripping, or the determination of the hydrogen underpotential deposition (Hupd) charge, are standard. For the determination of the Pt loading, a straightforward and cheap procedure using digestion in aqua regia with subsequent conversion of Pt(IV) to Pt(II) and UV-vis measurements is introduced.

Preparing and testing Pt/C fuel cell catalysts is subject to continuous discussion in the scientific community with respect to reproducibility and best practice. With the presented work, we intend to present a step-by-step tutorial to make and test Pt/C catalysts, which can serve as benchmark for novel catalyst systems.

用薄膜旋转圆盘电极法制备和测试燃料电池催化剂的研究

We present a step-by-step tutorial to prepare proton exchange membrane fuel cell (PEMFC) catalysts, consisting of Pt nanoparticles (NPs) supported on a ...

Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri

The killifish Nothobranchius furzeri is an emerging model organism in the field of ecotoxicology and its applicability in acute and chronic ecotoxicity testing has been demonstrated. Overall, the sensitivity of the species to toxic compounds is in the range with, or higher than, that of other model species.
This work describes protocols for acute, chronic, and multigenerational bioassays of single and combined stressor effects on N. furzeri. Due to its short maturation time and life-cycle, this vertebrate model allows the study of endpoints such as maturation time and fecundity within four months. Transgenerational full life-cycle exposure trials can be performed in as little as 8 months. Since this species produces eggs that are drought-resistant and remain viable for years, the on-site culture of the species is not needed but individuals can be recruited when required. The protocols are designed to measure life-history traits (mortality, growth, fecundity, weight) and critical thermal maximum.

Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri

In this work, we describe an acute, chronic and multigenerational bioassay to study the effects of single and combined stressors on the Turquoise killifish Nothobranchius furzeri. This protocol is designed to study life-history traits (mortality, growth, fecundity, weight) and critical thermal maximum.

绿松石鳉鱼的急性和慢性毒性测试协议Nothobranchius furzeri

The killifish Nothobranchius furzeri is an emerging model organism in the field of ecotoxicology and its applicability in acute and chronic ecotoxicity ...

Bioindication Testing of Stream Environment Suitability for Young Freshwater Pearl Mussels Using In Situ Exposure Methods

Knowledge of habitat suitability for freshwater mussels is an important step in the conservation of this endangered species group. We describe a protocol for performing in situ juvenile exposure tests within oligotrophic river catchments over one-month and three-month periods. Two methods (in both modifications) are presented to evaluate the juvenile growth and survival rate. The methods and modifications differ in value for the locality bioindication and each has its benefits as well as limitations. The sandy cage method works with a large set of individuals, but only some of the individuals are measured and the results are evaluated in bulk. In the mesh cage method, the individuals are kept and measured separately, but a low individual number is evaluated. The open water exposure modification is relatively easy to apply; it shows the juvenile growth potential of sites and can also be effective for water toxicity testing. The within-bed exposure modification needs a high workload but is closer to the conditions of a natural juvenile environment and it is better for reporting the real suitability of localities. On the other hand, more replications are needed in this modification due to its high-hyporheic environment variability.

Bioindication Testing of Stream Environment Suitability for Young Freshwater Pearl Mussels Using In Situ Exposure Methods

In situ bioindications enable determination of the suitability of an environment for endangered mussel species. We describe two methods based on the juvenile exposure of freshwater pearl mussels in cages to oligotrophic river habitats. Both methods are implemented in variants for open water and hyporheic water environments.

用原位曝光法测定淡水珍珠贻贝 Bioindication 的流环境适宜性

Knowledge of habitat suitability for freshwater mussels is an important step in the conservation of this endangered species group. We describe a protocol ...

Metal Corrosion and the Efficiency of Corrosion Inhibitors in Less Conductive Media

Material corrosion can be a limiting factor for different materials in many applications. Thus, it is necessary to better understand corrosion processes, prevent them and minimize the damages associated with them. One of the most important characteristics of corrosion processes is the corrosion rate. The measurement of corrosion rates is often very difficult or even impossible especially in less conductive, non-aqueous environments such as biofuels. Here, we present five different methods for the determination of corrosion rates and the efficiency of anti-corrosion protection in biofuels: (i) a static test, (ii) a dynamic test, (iii) a static test with a reflux cooler and electrochemical measurements (iv) in a two-electrode arrangement and (v) in a three-electrode arrangement. The static test is advantageous due to its low demands on material and instrumental equipment. The dynamic test allows for the testing of corrosion rates of metallic materials at more severe conditions. The static test with a reflux cooler allows for the testing in environments with higher viscosity (e.g., engine oils) at higher temperatures in the presence of oxidation or an inert atmosphere. The electrochemical measurements provide a more comprehensive view on corrosion processes. The presented cell geometries and arrangements (the two-electrode and three-electrode systems) make it possible to perform measurements in biofuel environments without base electrolytes that could have a negative impact on the results and load them with measurement errors. The presented methods make it possible to study the corrosion aggressiveness of an environment, the corrosion resistance of metallic materials, and the efficiency of corrosion inhibitors with representative and reproducible results. The results obtained using these methods can help to understand corrosion processes in more detail to minimize the damages caused by corrosion.

The testing of processes associated with material corrosion can often be difficult especially in non-aqueous environments. Here, we present different methods for short-term and long-term testing of corrosion behavior of non-aqueous environments such as biofuels, especially those containing bioethanol.

金属腐蚀与腐蚀抑制剂在低导电介质中的效率

Material corrosion can be a limiting factor for different materials in many applications. Thus, it is necessary to better understand corrosion processes, ...

A Small-Scale Setup for Algal Toxicity Testing of Nanomaterials and Other Difficult Substances

Ecotoxicity data is a requirement for pre- and post-market registration of chemicals by European and international regulations (e.g., REACH). The algal toxicity test is frequently used in regulatory risk assessment of chemicals. In order to achieve high reliability and reproducibility the development of standardized guidelines is vital. For algal toxicity testing, the guidelines require stable and uniform conditions of parameters such as pH, temperature, carbon dioxide levels and light intensity. Nanomaterials and other so-called difficult substances can interfere with light causing a large variation in results obtained hampering their regulatory acceptance. To address these challenges, we have developed LEVITATT (LED Vertical Illumination Table for Algal Toxicity Tests). The setup utilizes LED illumination from below allowing for a homogenous light distribution and temperature control while also minimizing intra-sample shading. The setup optimizes the sample volume for biomass quantification and does at the same time ensure a sufficient influx of CO2 to support exponential growth of the algae. Additionally, the material of the test containers can be tailored to minimize adsorption and volatilization. When testing colored substances or particle suspensions, the use of LED lights also allows for increasing the light intensity without additional heat generation. The compact design and minimal equipment requirements increase the possibilities for implementation of the LEVITATT in a wide range of laboratories. While compliant with standardized ISO and OECD guidelines for algal toxicity testing, LEVITATT also showed a lower inter-sample variability for two reference substances (3,5-Dicholorophenol and K2Cr2O7) and three nanomaterials (ZnO, CeO2, and BaSO4) compared to Erlenmeyer flasks and microtiter plates.

We demonstrate algal toxicity testing for difficult substances (e.g., colored substances or nanomaterials) using a setup illuminated vertically with an LED.

纳米材料和其他难物质藻类毒性试验的小型装置

Ecotoxicity data is a requirement for pre- and post-market registration of chemicals by European and international regulations (e.g., REACH). The algal ...