我们感谢赖斯大学在多孔介质 (休斯敦, TX, 美国) 过程中的资金支持。

注意: 本协议涉及处理高温烘箱、有毒化学品和 UV 光。请仔细阅读所有的材料安全数据表, 并遵循您的机构的化学品安全指南。
1. 设备设计
<ol><li>在 CAD 软件应用程序中设计掩。<ol>
<li>绘制一个3厘米长和0.5 厘米宽的矩形通道 (图 1b-右上)。</li>
		<li>创建一个表示多孔介质颗粒的封闭形状数组。 注意: 这些形状被称为柱子, 因为它们将在软光刻过程中成为三维结构。柱子的形状和大小应该是在数十微米的顺序, 并且有间距十到100微米。可以使用多个开机自检大小来创建异质性, 而在介质中可以保留剖面以模拟断开。</li>
		<li>绘制大约1/3 宽于多孔介质剖面的进风口通道。绘制一个从入口端口产生的通道作为一个排水管。</li>
		<li>在整个设计中绘制一个边界框, 从设计中至少要有1.0 厘米的间隙。 注意: 边界框和设计边框之间的区域, 以及柱子, 都要在掩上透明。</li>
	</ol>
</li>
	<li>向公司提交 cad 文件以进行高分辨率 cad 打印 注: 可选: 用于泡沫驱实验, 设计微流体泡沫发生器 (图 1a)。重复步骤 1, 省略了设计异质性和边界框。在多孔介质设计之前, 建议在入口处引入流聚焦几何。掩上的流动空间应该是透明的。</li>
</ol>2. 模具制造
<ol><li>在洁净室中创建光刻胶图案的硅片主模<ol>
<li>自旋涂层20µm 层的光刻胶到一个新的硅片在 2000 rpm 三十年代。</li>
		<li>软烘烤圆片在一个热的板材在二增量:65 ° c 为 1 min 跟随95° c 为 3 min。</li>
		<li>使用掩模器将光刻胶层与 CAD 设计一起使用恒定用量为150兆焦耳/cm2。</li>
		<li>在热板上进行两次后烘烤:65 ° c, 1 分钟, 其次为95° c, 3 分钟. 允许硅片冷却5分钟。</li>
		<li>在玻璃结晶盘中, 将硅片浸入100毫升的丙烯乙二醇甲基醚醋酸酯中。用手轻轻搅动10分钟以开发光刻胶图案。用异丙醇冲洗, 在干燥的气流下烘干晶片。</li>
		<li>硬烤晶圆片上的热板两个增量: 120 ° c 为5分钟, 其次为150° c 为 10 min. 允许硅片冷却15分钟。</li>
	</ol>
</li>
	<li>铸造硅晶片到硅片主模<ol>
<li>在无尘一次性容器内共混合30克的硅橡胶弹性体和固化剂, 其比例为5:1。</li>
		<li>在真空干燥的30分钟。</li>
		<li>在一个 150 mm 的玻璃培养皿中, 将硅晶片倒入光刻胶图案的硅片主模上。</li>
		<li>在80° c 烤箱中放置含有晶片和硅氧烷的培养皿1小时。</li>
		<li>从烤箱中取出培养皿, 使其含量达到室温。 注意: 此过程可能在此时暂停。</li>
	</ol>
</li>
	<li>将模式转移到光学粘合剂中的硅橡胶模具的制备<ol>
<li>小心地用手术刀切割出的硅氧烷模具, 并剥离模具远离晶片。</li>
		<li>使用透明胶带清洁和保护硅橡胶模具。 注意: 此过程可能在此时暂停。</li>
		<li>放置在一个无尘 60 mm 塑料培养皿底部的硅橡胶模具, 图案的一面。让十年代的硅橡胶坚持塑料。</li>
		<li>用透明胶带保护表面, 直到台阶3.1.1。 注: 可选: 制造泡沫发生器, 重复步骤2.1。通过2.3.2。用于泡沫发生器的设计。</li>
	</ol>
</li>
</ol>3. 光学粘接装置的制作
<ol><li>将光学粘合剂投射到硅橡胶模具上<ol>
<li>将胶带从图案表面取出, 并将光学粘合剂倒入 150 mm 培养皿中, 使其深度约0.9 厘米以上。用棉签轻轻地除去任何气泡。</li>
	</ol>
</li>
	<li>在紫外光系统中, 在紫外线照射下固化光粘合剂共40分钟, 如 3.2.1-3.2.5 步骤所述。 注意: 在使用 UV 光源时应佩戴适当的防护。<ol>
<li>将培养皿暴露在 UV 光 (254 nm) 中5分钟。</li>
		<li>反转培养皿, 使底部现在面对 uv 光源, 并将残缺不全暴露在紫外线下5分钟。</li>
		<li>反转培养皿, 将其返回至直立位置, 并将顶部重新至 UV 光5分钟。</li>
		<li>再次倒置的培养皿, 并重新底部的一面紫外线10分钟。</li>
		<li>将培养皿反转至直立位置, 并将顶部重新至 UV 光15分钟。 注: 只有在使用指定的 PSD-UV 设备 (材料表) 时, 3.2.1 通过3.2.5 的步骤中的固化过程才适用。固化时间会因所使用的特定光源和光学粘合层的精确厚度而异。</li>
	</ol>
</li>
	<li>从硅橡胶模具中取出固化的光学粘合剂<ol>
<li>使用盒式切割机, 仔细打破了培养皿模具的光学粘合剂。 注意: 框刀刀片非常锋利, 可以很容易地切割肉。在坏的培养皿的锋利边缘工作时要小心。</li>
		<li>使用一把坚固的剪刀从设计的边缘去除多余的光学粘合剂。</li>
		<li>慢慢剥离的硅橡胶模具远离光学胶粘剂冰球。用透明胶带保护光学胶粘剂表面的图案部分和硅橡胶表面。</li>
		<li>使用1.0 毫米活检冲床, 以创建入口, 出口和排水孔。用透明胶带保护带图案的光学粘合剂。</li>
	</ol>
</li>
	<li>准备基板<ol>
<li>将1毫升的光学粘合剂放到新的玻片上, 并在两个步骤中旋转滑动: 500 rpm 为5秒, 二十年代为 4000 rpm。</li>
		<li>将基板快速转移到 uv 光处理, 并在三十年代 uv 光下部分固化薄光胶层。</li>
	</ol>
</li>
	<li>粘合在基板上的光学粘合剂<ol>
<li>将光学粘合剂浇铸, 图案的一面上, 和承印物, 涂边, 在一个 O2等离子清洗器。等离子清洁表面为二十年代在 540 mTorr。</li>
		<li>将两个待处理的表面牢固地压紧, 直到所有不受欢迎的空气袋被最小化或移除。</li>
		<li>在紫外线照射下充分固化20分钟的设备。 注意: 对于 UV 灯, 请佩戴防护眼镜、实验室大衣、手套、等的适当保护。</li>
		<li>将设备放在50° c 的热板上18小时。</li>
	</ol>
</li>
	<li>将6英寸长的 0.58 mm ID 低密度聚乙烯油管 (PE/3) 插入设备上的每个端口。</li>
	<li>使用5分钟 quick-set 环氧树脂, 以确保油管到位。 注: 可选: 要完成泡沫发生器, 重复步骤 3.5.1, 3.5.2, 3.6, 和3.7。使用泡沫发生器和新的玻璃幻灯片, 而不是光学粘合剂铸造和准备基板, 分别。</li>
</ol>4. 驱油试验
<ol><li>在装有高速摄影机的倒置显微镜上, 准备微流控装置的影像。使用磁带将设备固定到显微镜阶段。使用4X 目标, 关注感兴趣的领域 (君)。</li>
	<li>准备注射液 注: 对于三相系统, 应将染料添加到清除置换液中, 以便为图像分析提供颜色对比度。<ol>
<li>将3毫升的原油或模型油样品装入10毫升的玻璃注射器中, 配有23计的工业配药尖端。将注射器固定在注射器泵座上, 并在注射器泵设置上设置合适的直径值。</li>
		<li>将1毫升的置换液装入3毫升的塑料注射器, 配有23计的工业配药尖端。将注射器固定在注射器泵座上, 并在注射器泵设置上设置合适的直径值。 注: 可选: 用于发泡试验, 将10米长的25µm 直径的玻璃毛细管连接到 N 个2气罐中, 并将气体压力设置为所需的气体流速, 以达到从校准曲线中获得的要求。允许10分钟的气体流量到平衡。</li>
	</ol>
</li>
	<li>含油的光学粘接型多孔介质装置的饱和<ol>
<li>通过将针尖插入 PE/3 管, 将置换液连接到装置的入口。 注: 可选: 当泡沫被用作置换阶段时, 将置换液注射器连接到泡沫发生器的入口。通过将毛细管插入23口径工业配药尖端, 并用 quick-set 环氧密封环缝, 将气毛细管连接到泡沫发生器的第二入口口。发泡发生器的出口, 然后使用23口径连接器连接到光学粘接装置的入口。</li>
		<li>通过将针头插入 PE/3 油管, 将油注射器连接到装置的入口。</li>
		<li>开始在2毫升/小时内将油进入光学粘接装置的出口口, 同时将置换液流动到0.8 毫升/小时的入口口, 这样两种流体都流出排泄口。置换液不应进入多孔介质。收集20毫升玻璃瓶中的废水。</li>
	</ol>
</li>
	<li>开始拍摄的多孔介质设备上的一帧速率足够快, 以捕捉所需的现象。一个典型的帧速率是 50 fps。捕获100% 油饱和区域的静止图像。</li>
	<li>迅速和同时削减 PE/3 油管是流动在油使用剪刀, 而夹在排水管管与一个5厘米的粘合剂钳。</li>
	<li>允许置换液侵入设备, 直到油排量达到稳定状态或相机耗尽内存。</li>
</ol>5. 图像和数据分析
<ol><li>使用免费的图像分析软件, 如图像 J, 或使用 MATLAB 中的图像分析工具箱对实验的素材进行分析。<ol>
<li>利用100% 油饱和通道的静止图像, 计算了多孔介质的孔隙率。</li>
	</ol>
</li>
	<li>用以下公式计算孔隙体积: <img alt="Equation 1" src="/files/ftp_upload/56592/56592eq1.jpg">
</li>
	<li>利用图像分析软件确定的油饱和度, 作为总流空间的一小部分, 在每个帧的视频片段中进行实验。对于两相位移实验, 每个框架的置换相饱和度可以计算为: <img alt="Equation 2" src="/files/ftp_upload/56592/56592eq2.jpg">
</li>
	<li>准备一个含油饱和度与注入液的孔隙量的图 注: 可选: 对于三相系统, 如泡沫位移实验, 使用 MATLAB 图像分析工具箱, 使用每个阶段的特征 RGB 范围对每个置换阶段进行颜色分类。准备一个图, 显示饱和的所有三阶段与注入孔隙体积。</li>
</ol>

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F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Norland+optical+adhesive+(NOA81)+microchannels+with+adjustable+wetting+behavior+and+high+chemical+resistance+against+a+range+of+mid-infrared-transparent+organic+solvents.">Norland optical adhesive (NOA81) microchannels with adjustable wetting behavior and high chemical resistance against a range of mid-infrared-transparent organic solvents.</a> Sensor Actuat B-Chem. 156 (2), 994-1001 (2011).</li><li>Hung, L. -. H., Lin, R., Lee, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+microfabrication+of+solvent-resistant+biocompatible+microfluidic+devices.">Rapid microfabrication of solvent-resistant biocompatible microfluidic devices.</a> Lab Chip. 8 (6), 983 (2008).</li></ol>

作者没有什么可透露的。

本协议研究光学胶粘剂中的油回收过程, 微观在 non-polymeric 微观 (如玻璃或硅) 的鲁棒性和微流体器件的简易制造之间取得了平衡。与玻璃或光学粘合剂制成的微观不同的是, 该设备缺乏对光有机物种的抵抗力。由于这些器件的表面具有不稳定的润湿性能, 而且聚合物基体可渗透到气体19, 因此, 微观也不太适合进行许多实验。相比之下, 光学胶粘剂的润湿性比硅橡胶更稳定, 而且对?...

该方法的目的是在多孔介质中对多相、多组分流体相互作用和复杂的孔隙尺度动力学进行可视化和分析。多孔介质中的流体流动和输送多年来一直引起关注, 因为这些系统适用于一些地下过程, 如石油回收、含水层修复和水力压裂1,2,3,4,5. 使用微观模拟这些复杂的孔隙结构, 通过可视化不同流体相之间的孔隙级动态事件和媒体6、7、8来获得独特的洞察力,9,10,11。
传统的硅微观的制造成本昂贵, 耗时, 而且具有挑战性, 但从光学粘合剂构建微观提供了一个相对便宜, 快速, 方便的替代12,13, 14,15。与其它聚合物微观相比, 光学胶粘剂具有更稳定的表面润湿性能。例如, 在典型的位移实验16过程中, 烷 (模型) 的表面会很快变得疏水性。此外, 该杨氏模量为 2.5 mpa, 而光学胶粘剂的弹性模量为325兆帕斯卡13,17,18。因此, 光学胶粘剂不易受压力引起的变形和通道失效。重要的是, 固化的光学胶粘剂是更抗膨胀的低分子量有机成分, 这使得实验涉及原油和轻溶剂进行18。整体而言, 光学胶粘剂是一种优越的替代品, 以硅的位移研究, 涉及原油时, 微观是令人望而却步的复杂或昂贵和高温和压力的研究是不需要的。
本出版物所述的协议提供了光学胶粘剂微观的 step-by 步骤制作说明, 并报告了确保在操作少量流体方面成功的微妙技巧。首先介绍了基于软光刻的光学胶粘剂的设计与制作微观。然后, 给出了流量控制的超低流速的流体置换策略。然后给出了一个典型的实验结果。本实验揭示了原油和异质多孔介质存在下泡沫失稳和传播行为。最后, 报道了典型的图像处理和数据分析。
本文所提供的方法适用于封闭微通道空间中涉及多相流和相互作用的可视化应用。具体地说, 该方法针对大于5和小于700µm 的特征微特征分辨率进行了优化. 典型流速为0.1 到1毫升/小时。在研究原油或光溶剂在水中或气态流体的位移时, 根据这些优化参数在环境条件下的顺序, 本协议应该是适当的。

<table>
	<tbody>
		<tr>
			<td>3 mL Leur-Lok Syringe</td>
			<td>Fischer Scientific</td>
			<td>14-823-435</td>
			<td></td>
		</tr>
		<tr>
			<td>10 mL Glass Syringe</td>
			<td>Fischer Scientific</td>
			<td>1482698G</td>
			<td></td>
		</tr>
		<tr>
			<td>Photomask</td>
			<td>CAD/Art Services</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Silicon Wafer</td>
			<td>University Wafer</td>
			<td>452</td>
			<td></td>
		</tr>
		<tr>
			<td>Propylene-Glycol-Methyl-Ether-Acetate&#160;</td>
			<td>Sigma Aldrich</td>
			<td>484431-4L</td>
			<td></td>
		</tr>
		<tr>
			<td>150 mm Glass Petri Dish</td>
			<td>Carolina Biological Supply</td>
			<td>#721134</td>
			<td></td>
		</tr>
		<tr>
			<td>60 mm Plastic Petri Dish</td>
			<td>Carolina Biological Supply</td>
			<td>#741246</td>
			<td></td>
		</tr>
		<tr>
			<td>Mask Aligner</td>
			<td>EV Group</td>
			<td>EVG 620</td>
			<td></td>
		</tr>
		<tr>
			<td>1 mm Biopsy Punch</td>
			<td>Miltex, Plainsboro, NJ</td>
			<td>69031-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Industrial Dispensing Tip</td>
			<td>CML Supply</td>
			<td>Gauge 23</td>
			<td></td>
		</tr>
		<tr>
			<td>Inverted Microscope</td>
			<td>Olympus</td>
			<td>IX-71</td>
			<td></td>
		</tr>
		<tr>
			<td>Plasma System</td>
			<td>Harrick Plasma</td>
			<td>PDC-32G</td>
			<td>Plasma cleaner</td>
		</tr>
		<tr>
			<td>Polydimehtylsiloxane (PDMS)</td>
			<td>Dow Corning, Midland, MI</td>
			<td>SYLGARD 184</td>
			<td></td>
		</tr>
		<tr>
			<td>Norland Optical Adhesive 81 (NOA81) or (OA)</td>
			<td>Norland Products Inc.</td>
			<td>8116</td>
			<td>Optical adhesive</td>
		</tr>
		<tr>
			<td>Quick-Set Epoxy</td>
			<td>Fisher Scientific</td>
			<td>4001</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass Slides</td>
			<td>Globe Scientic Inc.</td>
			<td>1321</td>
			<td></td>
		</tr>
		<tr>
			<td>SU-8 2015 Photoresist</td>
			<td>MicroChem</td>
			<td>SU-8 2015</td>
			<td>Photo resist</td>
		</tr>
		<tr>
			<td>Syringe Pump</td>
			<td>Harvard Apparatus</td>
			<td>Fusion 400</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass Capillary Tubing</td>
			<td>SGE Analytical Science</td>
			<td>1154710C</td>
			<td></td>
		</tr>
		<tr>
			<td>High-Speed Camera</td>
			<td>Vision Research</td>
			<td>V 4.3</td>
			<td></td>
		</tr>
		<tr>
			<td>Polyethylene Tubing</td>
			<td>Scientific Commodities Inc.</td>
			<td>#BB31695-PE/3</td>
			<td></td>
		</tr>
	</tbody>
</table>

microfluidic devices for characterizing pore-scale event processes in porous media for oil recovery applications

微流控设备是一种用于研究微观尺度的运输过程的多功能工具。不像传统的烷 (聚氧烷) 器件, 对低分子量的油成分具有抗微流控装置的需求。在这里, 我们演示了一种简便的方法, 使设备的这一属性, 我们使用该协议的产品, 以检查的孔隙规模机制, 泡沫恢复原油。模式首先使用计算机辅助设计 (CAD) 软件设计, 并在高分辨率打印机的透明度上打印。然后通过光刻过程将这种模式转移到光刻胶中。在图案上浇铸, 在烤箱中固化, 然后取下以获得霉菌。一种硫醇烯交联聚合物, 通常用作光学粘合剂 (OA), 然后浇在模具上, 在紫外线照射下固化。该模具被剥离的光学粘合剂铸造。然后准备一个玻璃基板, 并将该设备的两个半部分粘合在一起。光学 adhesive-based 器件比传统的硅橡胶微流控器件具有更强的鲁棒性。环氧树脂结构是耐膨胀的许多有机溶剂, 这为实验提供了新的可能性, 涉及轻有机液体。此外, 这些器件的表面润湿性比硅橡胶更稳定。光学粘接微流控装置的建设是简单的, 但需要更多的努力比 PDMS-based 设备的制造。此外, 虽然光学粘接装置在有机液体中是稳定的, 但在很长一段时间后, 它们可能会表现出较低的粘结强度。光学粘接微流体器件可以在几何中做为多孔介质的2维微观。这些装置被应用于研究石油驱替, 以提高我们对提高石油采收率和含水层修复所涉及的孔隙规模机制的认识。

在这个例子实验中, 水沫被用来取代中东原油 (具有 5.4 cP 的黏度和40°的 API 重力) 在具有层状渗透率对比度的非均匀多孔介质中。一种模型的泡沫发生器连接到以前完全浸透原油的光学粘合剂。图 1a显示了掩泡沫发生器、光刻胶图案硅晶片和已完成的泡沫发生器的 CAD 设计, 并插入了入口和出口管。图 1b显示了具有分层渗透率对?...

Watch this Scientific Journal Video about Microfluidic Devices for Characterizing Pore-scale Event Processes in Porous Media for Oil Recovery Applications at JoVE.com

Microfluidic Devices for Characterizing Pore-scale Event Processes in Porous Media for Oil Recovery Applications

这一过程的目的是容易和迅速地产生一个可定制的几何和耐膨胀的有机流体的微流控油回收研究。首先生成一个烷模具, 然后用它来铸造氧设备。有代表性的位移研究报告。

微流体器件用于表征多孔介质中的孔隙尺度事件过程, 用于采油应用

Microfluidic devices are versatile tools for studying transport processes at a microscopic scale. A demand exists for microfluidic devices that are ...

microfluidic-devices-for-characterizing-pore-scale-event-processes

Research

JoVE Journal

Environment

10.3K 次观看.  Rice University. 这一过程的目的是容易和迅速地产生一个可定制的几何和耐膨胀的有机流体的微流控油回收研究。首先生成一个烷模具, 然后用它来铸造氧设备。有代表性的位移研究报告。

视频: Microfluidic Devices for Characterizing Pore-scale Event Processes in Porous Media for Oil Recovery Applications

Measurement of H2S in Crude Oil and Crude Oil Headspace Using Multidimensional Gas Chromatography, Deans Switching and Sulfur-selective Detection

A method for the analysis of dissolved hydrogen sulfide in crude oil samples is demonstrated using gas chromatography. In order to effectively eliminate interferences, a two dimensional column configuration is used, with a Deans switch employed to transfer hydrogen sulfide from the first to the second column (heart-cutting). Liquid crude samples are first separated on a dimethylpolysiloxane column, and light gases are heart-cut and further separated on a bonded porous layer open tubular (PLOT) column that is able to separate hydrogen sulfide from other light sulfur species. Hydrogen sulfide is then detected with a sulfur chemiluminescence detector, adding an additional layer of selectivity. Following separation and detection of hydrogen sulfide, the system is backflushed to remove the high-boiling hydrocarbons present in the crude samples and to preserve chromatographic integrity. Dissolved hydrogen sulfide has been quantified in liquid samples from 1.1 to 500 ppm, demonstrating wide applicability to a range of samples. The method has also been successfully applied for the analysis of gas samples from crude oil headspace and process gas bags, with measurement from 0.7 to 9,700 ppm hydrogen sulfide.

Measurement of H2S in Crude Oil and Crude Oil Headspace Using Multidimensional Gas Chromatography, Deans Switching and Sulfur-selective Detection

A multidimensional gas chromatography method for the analysis of dissolved hydrogen sulfide in liquid crude oil samples is presented. A Deans switch is used to heart-cut light sulfur gases for separation on a secondary column and detection on a sulfur chemiluminescence detector.

h的测量<sub&gt; 2</sub&gt; S在原油和原油顶空用多维气相色谱法，院长交换和选择性硫检测

A method for the analysis of dissolved hydrogen sulfide in crude oil samples is demonstrated using gas chromatography. In order to effectively eliminate ...

科研

环境科学

A Gusseted Thermogradient Table to Control Soil Temperatures for Evaluating Plant Growth and Monitoring Soil Processes

Thermogradient tables were first developed in the 1950s primarily to test seed germination over a range of temperatures simultaneously without using a series of incubators. A temperature gradient is passively established across the surface of the table between the heated and cooled ends and is lost quickly at distances above the surface. Since temperature is only controlled on the table surface, experiments are restricted to shallow containers, such as Petri dishes, placed on the table. Welding continuous aluminum vertical strips or gussets perpendicular to the surface of a table enables temperature control in depth via convective heat flow. Soil in the channels between gussets was maintained across a gradient of temperatures allowing a greater diversity of experimentation. The gusseted design was evaluated by germinating oat, lettuce, tomato, and melon seeds. Soil temperatures were monitored using individual, battery-powered dataloggers positioned across the table. LED lights installed in the lids or along the sides of the gradient table create a controlled temperature chamber where seedlings can be grown over a range of temperatures. The gusseted design enabled accurate determination of optimum temperatures for fastest germination rate and the highest percentage germination for each species. Germination information from gradient table experiments can help predict seed germination and seedling growth under the adverse soil conditions often encountered during field crop production. Temperature effects on seed germination, seedling growth, and soil ecology can be tested under controlled conditions in a laboratory using a gusseted thermogradient table.

Traditional thermogradient tables create a range of temperatures across the surface. Welding gussets perpendicular to the surface of a thermogradient table will control temperature in depth increasing possible research applications.

一个插角Thermogradient表来控制土壤温度为评估植物生长和监测土壤过程

Thermogradient tables were first developed in the 1950s primarily to test seed germination over a range of temperatures simultaneously without using a ...

A Lipid Extraction and Analysis Method for Characterizing Soil Microbes in Experiments with Many Samples

Microbial communities are important drivers and regulators of ecosystem processes. To understand how management of ecosystems may affect microbial communities, a relatively precise but effort-intensive technique to assay microbial community composition is phospholipid fatty acid (PLFA) analysis. PLFA was developed to analyze phospholipid biomarkers, which can be used as indicators of microbial biomass and the composition of broad functional groups of fungi and bacteria. It has commonly been used to compare soils under alternative plant communities, ecology, and management regimes. The PLFA method has been shown to be sensitive to detecting shifts in microbial community composition.
An alternative method, fatty acid methyl ester extraction and analysis (MIDI-FA) was developed for rapid extraction of total lipids, without separation of the phospholipid fraction, from pure cultures as a microbial identification technique. This method is rapid but is less suited for soil samples because it lacks an initial step separating soil particles and begins instead with a saponification reaction that likely produces artifacts from the background organic matter in the soil. 
This article describes a method that increases throughput while balancing effort and accuracy for extraction of lipids from the cell membranes of microorganisms for use in characterizing both total lipids and the relative abundance of indicator lipids to determine soil microbial community structure in studies with many samples. The method combines the accuracy achieved through PLFA profiling by extracting and concentrating soil lipids as a first step, and a reduction in effort by saponifying the organic material extracted and processing with the MIDI-FA method as a second step.

The article describes a method that increases throughput while balancing effort and accuracy for extraction of lipids from the cell membranes of microorganisms for use in characterizing both total lipids and the relative abundance of indicator lipids to determine soil microbial community structure in studies with many samples.

用于表征土壤微生物的脂质提取和分析方法在许多样品的实验中

Microbial communities are important drivers and regulators of ecosystem processes. To understand how management of ecosystems may affect microbial ...

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 ...

Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

Dimethylated thioarsenicals such as dimethylmonothioarsinic acid (DMMTAV) and dimethyldithioarsinic acid (DMDTAV), which are produced by the metabolic pathway of dimethylarsinic acid (DMAV) thiolation, have been recently found in the environment as well as human organs. DMMTAV and DMDTAV can be quantified to determine the ecological effects of dimethylated thioarsenicals and their stability in environmental media. The synthesis method for these compounds is unstandardized, making replicating previous studies challenging. Furthermore, there is a lack of information about storage techniques, including storage of compounds without species transformation. Moreover, because only limited information about synthesis methods is available, there may be experimental difficulties in synthesizing standard chemicals and performing quantitative analysis. The protocol presented herein provides a practically modified synthesis method for the dimethylated thioarsenicals, DMMTAV and DMDTAV, and will help in the quantification of species separation analysis using high performance liquid chromatography in conjunction with inductively coupled plasma mass spectrometry (HPLC-ICP-MS). The experimental steps of this procedure were modified by focusing on the preparation of chemical reagents, filtration methods, and storage.

Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

This article presents modified experimental protocols for dimethylmonothioarsinic acid (DMMTAV) and dimethyldithioarsinic acid (DMDTAV) synthesis, inducing dimethylarsinic acid (DMAV) thiolation through mixing of DMAV, Na2S, and H2SO4. The modified protocol provides an experimental guideline, thereby overcoming limitations of the synthesis steps that could have caused experimental failures in quantitative analysis.

使用 DMAv为环境应用 DMMTAv和 DMDTAv的准备: 合成、纯化和确认

Dimethylated thioarsenicals such as dimethylmonothioarsinic acid (DMMTAV) and dimethyldithioarsinic acid (DMDTAV), which are produced by the metabolic ...

Experimental Study of the Relationship Between Particle Size and Methane Sorption Capacity in Shale

The amount of adsorbed shale gas is a key parameter used in shale gas resource evaluation and target area selection, and it is also an important standard for evaluating the mining value of shale gas. Currently, studies on the correlation between particle size and methane adsorption are controversial. In this study, an isothermal adsorption apparatus, the gravimetric sorption analyzer, is used to test the adsorption capacity of different particle sizes in shale to determine the relationship between the particle size and the adsorption capacity of shale. Thegravimetric method requires fewer parameters and produces better results in terms of accuracy and consistency than methods like the volumetric method. Gravimetric measurements are performed in four steps: a blank measurement, preprocessing, a buoyancy measurement, and adsorption and desorption measurements. Gravimetric measurement is presently considered to be a more scientific and accurate method of measuring the amount of adsorption; however, it is time-consuming and requires a strict measuring technique. A Magnetic Suspension Balance (MSB) is the key to verify the accuracy and consistency of this method. Our results show that adsorption capacity and particle size are correlated, but not a linear correlation, and the adsorptions in particles sieved into 40 - 60 and 60 - 80 meshes tend to be larger. We propose that the maximum adsorption corresponding to the particle size is approximately 250 &#181;m (60 mesh) in the shale gas fracturing.

We use an isothermal adsorption apparatus, the gravimetric sorption analyzer, to test the adsorption capacity of different particle sizes of shale, in order to find out the relationship between particle size and the adsorption capacity of shale.

页岩中粒径与甲烷吸附能力关系的实验研究

The amount of adsorbed shale gas is a key parameter used in shale gas resource evaluation and target area selection, and it is also an important standard ...

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, ...

Characterizing Microbiome Dynamics &#8211; Flow Cytometry Based Workflows from Pure Cultures to Natural Communities

The investigation of pure cultures and monitoring of microbial community dynamics is vital to understand and control natural ecosystems and technical applications driven by microorganisms. Next generation sequencing methods are widely utilized to resolve microbiomes, but they are generally resource and time intensive and deliver mostly qualitative information. Flow cytometric microbiome analysis does not suffer from those disadvantages and can provide relative subcommunity abundances and absolute cell numbers at-line. Although it does not deliver direct phylogenetic information, it can enhance the analysis depth and resolution of sequencing approaches. In sharp contrast to medical applications in both research and routine settings, flow cytometry is still not widely used for microbiome analysis. Missing information on sample preparation and data analysis pipelines may create an entry barrier for the researchers facing microbiome analysis challenges that would often be textbook flow cytometry applications. Here, we present three comprehensive workflows for pure cultures, complex communities in clear medium and complex communities in challenging matrices, respectively. We describe individual sampling and fixation procedures and optimized staining protocols for the respective sample sets. We elaborate the cytometric analysis with a complex research centered and an application focused bench top device, describe the cell sorting procedure and suggest data analysis packages. We furthermore propose important experimental controls and apply the presented workflows to the respective sample sets.

Flow cytometric analysis has proven valuable for investigating pure cultures and monitoring microbial community dynamics. We present three comprehensive workflows, from sampling to data analysis, for pure cultures and complex communities in clear medium as well as in challenging matrices, respectively.

微生物动力学的特征-基于流式细胞仪的工作流从纯文化到自然群落

The investigation of pure cultures and monitoring of microbial community dynamics is vital to understand and control natural ecosystems and technical ...

Single-throughput Complementary High-resolution Analytical Techniques for Characterizing Complex Natural Organic Matter Mixtures

Natural organic matter (NOM) is composed of a highly complex mixture of thousands of organic compounds which, historically, proved difficult to characterize. However, to understand the thermodynamic and kinetic controls on greenhouse gas (carbon dioxide [CO2] and methane [CH4]) production resulting from the decomposition of NOM, a molecular-level characterization coupled with microbial proteome analyses is necessary. Further, climate and environmental changes are expected to perturb natural ecosystems, potentially upsetting complex interactions that influence both the supply of organic matter substrates and the microorganisms performing the transformations. A detailed molecular characterization of the organic matter, microbial proteomics, and the pathways and transformations by which organic matter is decomposed will be necessary to predict the direction and magnitude of the effects of environmental changes. This article describes a methodological throughput for comprehensive metabolite characterization in a single sample by direct injection Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS), gas chromatography mass spectrometry (GC-MS), nuclear magnetic resonance (NMR) spectroscopy, liquid chromatography mass spectrometry (LC-MS), and proteomics analysis. This approach results in a fully-paired dataset which improves statistical confidence for inferring pathways of organic matter decomposition, the resulting CO2 and CH4 production rates, and their responses to environmental perturbation. Herein we present results of applying this method to NOM samples collected from peatlands; however, the protocol is applicable to any NOM sample (e.g., peat, forested soils, marine sediments, etc.).

This protocol describes a single throughput for complementary analytical and omics techniques culminating in a fully-paired characterization of natural organic matter and microbial proteomics in different ecosystems. This approach permits robust comparisons for identifying metabolic pathways and transformations important for describing greenhouse gas production and predicting responses to environmental change.

用于表征复杂天然有机物混合物的单吞吐量互补高分辨率分析技术

Natural organic matter (NOM) is composed of a highly complex mixture of thousands of organic compounds which, historically, proved difficult to ...

Combining Fluidic Devices with Microscopy and Flow Cytometry to Study Microbial Transport in Porous Media Across Spatial Scales

Understanding the transport, dispersion and deposition of microorganisms in porous media is a complex scientific task comprising topics as diverse as hydrodynamics, ecology and environmental engineering. Modeling bacterial transport in porous environments at different spatial scales is critical to better predict the consequences of bacterial transport, yet current models often fail to up-scale from laboratory to field conditions. Here, we introduce experimental tools to study bacterial transport in porous media at two spatial scales. The aim of these tools is to obtain macroscopic observables (such as breakthrough curves or deposition profiles) of bacteria injected into transparent porous matrices. At the small scale (10-1000 &#181;m), microfluidic devices are combined with optical video-microscopy and image processing to obtain breakthrough curves and, at the same time, to track individual bacterial cells at the pore scale. At larger scale, flow cytometry is combined with a self-made robotic dispenser to obtain breakthrough curves. We illustrate the utility of these tools to better understand how bacteria are transported in complex porous media such as the hyporheic zone of streams. As these tools provide simultaneous measurements across scales, they pave the way for mechanism-based models, critically important for upscaling. Application of these tools may not only contribute to the development of novel bioremediation applications but also shed new light on the ecological strategies of microorganisms colonizing porous substrates.

Breakthrough curves (BTCs) are efficient tools to study the transport of bacteria in porous media. Here we introduce tools based on fluidic devices in combination with microscopy and flow cytometric counting to obtain BTCs.

将流体设备与显微镜和流式细胞学相结合，研究跨空间尺度的多孔介质中的微生物传输

Understanding the transport, dispersion and deposition of microorganisms in porous media is a complex scientific task comprising topics as diverse as ...

微流体器件用于表征多孔介质中的孔隙尺度事件过程, 用于采油应用

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