作者感谢梅拉妮·弗兰克皮娅Griesheimer，杰西卡·亨里奇，佩特拉詹克，杰西卡·麦尔和诺伯特Sickinger的技术和分析支持这项工作。 该项目BioBoost内提供的财政支持大大确认。 BioBoost是欧洲的一个研发项目由欧盟委员会（www.bioboost.eu）根据合同282873联合资助的第七框架计划内。

1.启动<ol><li>通过启动辅助的 N 2供给和裂解气风扇激活全热解和冷凝系统。冲洗待机时用氮气的500升小时-1热解试验台。通过在流程控制开风扇的菜单并调节其标称体积流量，使得反应器中的压力为3-8毫巴高于环境压力调节风扇。 注意:特别是在启动时，存在爆炸性空气积聚的风险增加。该系统必须是为了降低这种风险完全惰性的。 </li><li>填充生物油周期（ 即富含有机质缩合物）与乙二醇作为猝灭系统起始培养基的合适量，以允许所述泵和匀浆器的安全操作（ 例如 ，15个公斤的例子给出）。记录的起始材料的重量。 </li><li>用适当填写的Amoun水冷凝循环的水净化，以允许所述泵的安全操作（ 例如7公斤实例中所给出）。记录起始材料的重量。 </li><li>升温系统，包括热载体加热和所有辅助加热器，通过过程控制打开他们的菜单和输入想要的值（ 如 500℃左右）。辅助加热建议在反应器本身和连接管到，以防止蒸汽不受控制的缩合第一冷凝器。 </li><li>通过在冷却器开关启动两个凝结循环热交换器的冷却循环。 </li><li>在过程控制打开他们的菜单启动这两个冷凝循环泵，并点击激活。使用相同的菜单调整质量流量，以提供足够的冷却能力。例如，在约350公斤hr的速率再循环生物油-1和喷射入骤冷容器之前冷却下来至80℃。再循环水孔德nsate在约600公斤小时-1的速率，此外，供应至300kg hr的速率冷却水-1在8℃。 </li><li>交换机上的静电除尘器。 </li><li>后，两个凝结循环已经运行10-20分钟，检查淬火系统的喷嘴阻止和删除任何堵塞存在。 </li><li>通过打开斗式提升机和在过程控制热载体馈送螺杆的菜单启动热载体循环，并点击激活。置，以便允许由占该热分解反应的热需求的平滑的启动时的热载体的温度高于期望的反应器温度的值。例如，在开始生物质进料之前在操作期间千公斤hr的质量流量-1在520℃的温度，但加热至545℃，供给载热体。 注意:确保反应器的双螺杆一旦被热载体喂养自动启动螺钉被激活。否则有阻塞，甚至到馈送系统损坏的危险。 </li><li>系统之后（ 即，所有的温度）达到设定值，开始通过填充的生物质储存与所需的原料进料生物质。随后，打开锁料斗，并通过点击激活他们的菜单中的过程控制启动生物送料螺杆。慢慢增加进料速率，以防止过度的压力波动。 </li></ol> 2.步骤和观察操作过程中需要不断<ol><li>记录的生物量，以占平衡，并采取适当的样品供给量。 </li><li>检查所需的反应器温度（热载体的出口温度），并相应地调节热量载体循环的加热。 </li><li>通过调整其标称体积流量，以保持所需的反应器压力调节风扇。 </li><li>检查在喷嘴阻塞淬火系统的（下降质量流量和/或在淬火温度增加）。 </li><li>观察，以便及时发现过度缩放及早跨越气旋和淬火系统的压降。安装适当的措施，能够在操作过程中除去过量的比例，特别是在热解蒸气的第一温度降（通常是淬火系统的入口）的点。 <ol><li>例如，通过使用杆以除去机械缩放清洁管的横截面。密封杆与一个垫圈以防止空气摄入到淬火系统。在杆的入口点安装一个球阀，以进一步减少空气泄漏，如果该清洁不运转。 注意:通过插入杆清洁淬火系统的入口导致从反应器中的气体除去的临时堵塞。如果不能得到保证，该清洁在&lt;10秒进行的生物质进料必须停止。 </li></ol></li><li>同时监测凝结循环的冷凝温度并在必要时调整温度的过程中温控器设定点。 </li><li>从周期尽快除去冷凝为最大允许填充水平的80％，已达到（取决于缓冲罐的尺寸和供给的生物质的量和类型）。 </li><li>进行气相测量。测量的气体的量以及它的组合物（见在步骤4.5详情）。 注:主要的气态化合物包括N 2，CO，CO 2，CH 4，O 2和H 2。是可以预期的其它化合物，如C 2 H 4，C 2 H 6和C 3 H 8。一个气体测量系统的一个实例描述如下（参见步骤4.5）。 注意:如果该热解单元的部分在压力下操作时，空气的泄漏可能导致爆炸性环境的发展。强烈建议密切OB服务于裂解气中的氧气量。 </li></ol> 3.关闭<ol><li>要停止实验，只需关闭生物饲料和调节风扇，以保持所需的反应器压力。 </li><li>允许系统（热载体循环和冷凝循环），为另一个30-40分钟运行，以确保所有的剩余物热解和产品回收。 </li><li>关闭热载体循环的加热。 </li><li>同时关闭缩合周期和静电除尘器的泵。 </li><li>空既冷凝循环，并记录每个冷凝物的重量计。减去原料（见步骤1.2和1.3）建立余额之前的金额。 </li><li>允许焦炭收集容器冷却至室温，在惰性气氛中。权衡炭的量。 注意:焦炭可能会出现自燃的特性，和具体的护理应处理该材料时服用。 </li><li>克丽n中的生物油循环用新鲜的乙二醇，并用1水性冷凝循环:水和乙醇混合物。用适量填写（见步骤1.2和1.3），并允许30-40分钟运行。 </li></ol> 4.需要分析的设置&#39;干&#39;和&#39;元素碳&#39;余额<ol><li>执行以下原料分析（对适用的标准例子括号内）: <ol><li>确定水含量17。 </li><li>确定灰分含量18。 </li><li>确定的元素碳，氢和氮含量19。 注:强烈建议中的水含量分析每个实验日，因为在天气条件的差异可能影响原料的水分含量。取决于很多的尺寸，需要几个样品可靠地表征原料。更多的分析，如纤维分析和较高的热值是推荐的，但不是强制性用于设置上述余额。 </li></ol></li><li>执行以下焦炭粉的分析（适用标准的例子在参考文献中给出）: <ol><li>确定水含量17。 </li><li>确定灰分含量18。 </li><li>确定的元素碳，氢和氮含量19。 注意:假定离开用于设置余额的处理时炭不具有的水分含量。分析的过程中，可能会发生水分卷取，并且是所必需的其他两个分析的校正的水含量。 </li></ol></li><li>执行以下生物油的分析（适用的标准或其他推荐方法的例子括号内）: <ol><li>根据标准方法确定由体积卡尔 - 费歇尔滴定法中的水含量。溶解在无水甲醇中的样品和与碱的混合物滴定它，SO 2和的I 2的已知浓度（的材料的详细实施例在材料列表中给出）。水每摩尔与I 2一摩尔反应。 </li><li>通过采取FPBO的3040克样品测定的固体含量并在甲醇中溶解至约100毫升的最终溶液体积。在室温下搅拌10分钟的溶液中。过滤通过纤维素过滤器在2.5微米颗粒保留在溶液中，直至获得澄清的滤液用甲醇彻底冲洗残余物。干燥在105℃将固体残余物​​过夜，并确定剩余的权重。 </li><li>确定的元素碳，氢和氮含量19。 </li><li>根据标准方法确定由1 H NMR分析的乙二醇含量。溶解在氘代甲醇中，用3-（三甲硅烷基）中的溶液的FPBO样品丙酸-2,2,3,3- -d 4中酸钠盐（TMSP）作为参考材料（约0.1克FPBO 0.8克溶液）。例如，该溶液可含有44克甲醇和0.1g TMSP。离心溶解样品以除去固体。通过1 H核磁共振谱（NMR）分析样品。乙二醇的羟基显示在3.55-3.65 ppm的峰。 TMSP的参考峰周围出现0ppm的，并用于量化乙二醇含量。 注:启动与纯乙二醇导致在第一冷凝器的冷凝物的稀释液。这需要在质量和能量平衡的计算和对结果的呈现中加以考虑。人们非常希望，以确定个别的化学化合物。这样的分析方法是非常复杂的，由于广大不同的化合物和冷凝基质的性质。这种分析的描述是本文的讨论范围之内。还应当指出，上述分析仅仅需要用于设置余额和不足以用于说明生物油作为产物。涵盖FPBO应用标准正在筹备之中。 </li></ol></li><li>执行以下凝水分析（适用的标准例子括号内）: <ol><li>确定由卡尔·费休滴定法的水分含量（参见4.3.1节）。 </li><li>确定总有机碳作为非吹出有机碳20。 注:启动用纯水引线在第二冷凝器的冷凝液的稀释。这需要在质量和能量平衡的计算和对结果的呈现中加以考虑。 </li></ol></li><li>监测整个实验中的气体组成，因为组合物随时间的变化相当大。例如，在一个过程气相色谱每隔30-60分钟这里介绍的实验中分析了的产物气体。分析以下气体种类:东北，H 2，CO，CO 2，N 2，O 2，CH 4，和烷烃/烯烃的 C 2 -C 5的组件。 <ol><li>注入氖的恒定气体流量进入反应器中作为参考。计算基于参考体积流量，平均气体组成比，在实验的持续时间和物种的密度各气体物质的质量。为了确定热分解气体中的水含量，假定在最后的冷凝器的出口温度饱和的条件。 </li></ol></li></ol>

<ol>
	<li>Dahmen, N., Henrich, E., Dinjus, E., Weirich, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+bioliq+bioslurry+gasification+process+for+the+production+of+biosynfuels,+organic+chemicals,+and+energy.">The bioliq bioslurry gasification process for the production of biosynfuels, organic chemicals, and energy.</a> Energ. Sust. Soc. 2 (1), 1-44 (2012).</li><li>Ahmad, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zeolite-based+bifunctional+catalysts+for+the+single+step+synthesis+of+dimethyl+ether+from+CO-rich+synthesis+gas.">Zeolite-based bifunctional catalysts for the single step synthesis of dimethyl ether from CO-rich synthesis gas.</a> Fuel Process Technol. 121, 38-46 (2014).</li><li>Haro, P., Trippe, F., Stahl, R., Henrich, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bio-syngas+to+gasoline+and+olefins+via+DME+-+A+comprehensive+techno-economic+assessment.">Bio-syngas to gasoline and olefins via DME - A comprehensive techno-economic assessment.</a> App Energy. , (2013).</li><li>Henrich, E., Dahmen, N., Dinjus, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cost+estimate+for+biosynfuel+production+via+biosyncrude+gasification.">Cost estimate for biosynfuel production via biosyncrude gasification.</a> Biofuels, Bioprod. Bioref. 3, 28-41 (2009).</li><li>Zhang, X., Kumar, A., Arnold, U., Sauer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biomass-derived+oxymethylene+ethers+as+diesel+additives:+A+thermodynamic+analysis.">Biomass-derived oxymethylene ethers as diesel additives: A thermodynamic analysis.</a> Energ. Procedia. 61, 1921-1924 (2014).</li><li>Bridgwater, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Renewable+fuels+and+chemicals+by+thermal+processing+of+biomass.">Renewable fuels and chemicals by thermal processing of biomass.</a> Chem. Eng. J. 91, 87-102 (2003).</li><li>Meier, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=State-of-the-art+of+fast+pyrolysis+in+IEA+bioenergy+member+countries.">State-of-the-art of fast pyrolysis in IEA bioenergy member countries.</a> Renew. Sust. Energ. Rev. 20, 619-641 (2013).</li><li>Rammler, R., Weiss, H. J., Bu&#223;mann, A., Simo, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gewinnung+von+&#214;l+durch+Schwelen+von+&#214;lschiefer+und+Teersand+als+Beitrag+zur+Energieversorgung.">Gewinnung von &#214;l durch Schwelen von &#214;lschiefer und Teersand als Beitrag zur Energieversorgung.</a> Chem. Ing. Tech. 53, 96-104 (1981).</li><li>Tr&#246;ger, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Utilization+of+biogenic+residues+and+wastes+in+thermochemical+systems+for+the+production+of+fuels:+current+status+of+the+project.">Utilization of biogenic residues and wastes in thermochemical systems for the production of fuels: current status of the project.</a> Biofuels, Bioprod. Bioref. 7, 12-23 (2013).</li><li>Tr&#246;ger, N., Richter, D., Stahl, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+feedstock+composition+on+product+yields+and+energy+recovery+rates+of+fast+pyrolysis+products+from+different+straw+types.">Effect of feedstock composition on product yields and energy recovery rates of fast pyrolysis products from different straw types.</a> J. Anal. Appl. Pyr. 100, 158-165 (2013).</li><li>Henrich, E., Dahmen, N., Weirich, F., Reimert, R., Kornmayer, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+pyrolysis+of+lignocelluloses+in+a+twin+screw+mixer+reactor.">Fast pyrolysis of lignocelluloses in a twin screw mixer reactor.</a> Fuel Process Technol. 143, 151-161 (2016).</li><li>Dahmen, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=State+of+the+art+of+the+bioliq+process+for+synthetic+biofuels+production.">State of the art of the bioliq process for synthetic biofuels production.</a> Env. Prog. Sust. Energ. 31, 176-181 (2012).</li><li>Funke, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+pyrolysis+char+-+Assessment+of+alternative+uses+within+the+bioliq+concept.">Fast pyrolysis char - Assessment of alternative uses within the bioliq concept.</a> Bioresour. Technol. 200, 905-913 (2016).</li><li>Lehto, J., Oasmaa, A., Solantausta, Y., Kyt&#246;, M., Chiaramonti, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Fuel oil quality and combustion of fast pyrolysis bio-oils. , (2013).</li><li>Fahmi, R., Bridgwater, A. V., Donnison, I., Yates, N., Jones, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+lignin+and+inorganic+species+in+biomass+on+pyrolysis+oil+yields,+quality+and+stability.">The effect of lignin and inorganic species in biomass on pyrolysis oil yields, quality and stability.</a> Fuel. 87, 1230-1240 (2008).</li><li>Oasmaa, A., Solantausta, Y., Arpiainen, V., Kuoppala, E., Sipil&#228;, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+Pyrolysis+Bio-Oils+from+Wood+and+Agricultural+Residues.">Fast Pyrolysis Bio-Oils from Wood and Agricultural Residues.</a> Energ. &amp; Fuels. 24, 1380-1388 (2010).</li><li>DIN German Institute for Standardization. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> DIN EN ISO 18134-3 Solid biofuels - Determination of moisture content - Oven dry method - Part 3: Moisture in general analysis sample. , (2015).</li><li>DIN German Institute for Standardization. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> DIN EN ISO 18122 Solid biofuels - Determination of ash content. , (2016).</li><li>DIN German Institute for Standardization. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Institute for Standardization. DIN EN ISO 16948 Solid biofuels - Determination of total content of carbon, hydrogen and nitrogen. , (2015).</li><li>DIN German Institute for Standardization. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Institute for Standardization. DIN EN 1484 Water analysis - Guidelines for the determination of total organic carbon (TOC) and dissolved organic carbon (DOC). , (1997).</li><li>DIN German Institute for Standardization. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> ESO 16993: Solid biofuels - Conversion of analytical results from one basis to another. , (2015).</li><li>Bridgwater, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review+of+fast+pyrolysis+of+biomass+and+product+upgrading).">Review of fast pyrolysis of biomass and product upgrading).</a> Biomass Bioenerg. 38, 68-94 (2012).</li><li>Westerhof, R. J. M., Kuipers, N. J. M., Kersten, S. R. A., van Swaaij, W. P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlling+the+water+content+of+biomass+fast+pyrolysis+oil.">Controlling the water content of biomass fast pyrolysis oil.</a> Ind. Eng. Chem. Res. 46, 9238-9247 (2007).</li></ol>

The authors have nothing to disclose.

对于所有的实验中，工艺条件如原料的大小，进料速率，压力，反应温度，冷凝温度，和流速都热载体和冷凝液循环的是相同的。自然地，所定义的限度内变化，不能避免。为测试植物如这里介绍的工艺开发部，波动的可接受范围和操作的所需时间为可再现的实验需要计算和/或根据经验确定。例如，反应器温度，它是由热载流子离开反应器的温度指示，被控制为35℃以上从反应开始在全生物质的能力的反应的整个过程的标准偏差停止喂食生物质（通常约4小时）。在反应器中的压力被控制以300-500 Pa的标准偏差。在压力峰可能是由于fluc发生tuations生物质喂养。它建议以调节进料螺杆系统所考虑的生物量材料，以减少这种波动和确保恒定生物质流量。分别在第一和第二冷凝器的冷凝温度保持在3℃的标准偏差和1℃。 它应该在这一点提出的所有实验在相同的反应器温度（500℃）下进行了注明。这个温度并不一定反映其存在于每个特定原料22的最佳快速热解的温度。反应器的温度的变化可能会导致一个优化的热分解温度以更高的有机油产量。 均衡方法的选择是不平凡的生物质的快速热解，施加分馏冷凝和利用生物质具有高灰分时尤其如此。三种不同类型的balancin的摹已在上一节被提出。报告产品部分的收益率的"为获得"基础上，对实际问题有利，如设备和存储容量的设计，因为它报告给所预期的实际产品分布。但是，这些值由原料中的水和灰分含量遮蔽。尤其是对废弃生物质- 例如，秸秆，林业和修剪残留和生物&#39;三废&#39; -这是一个问题，因为这些原料具有广泛的水和无机内容见表1。 生物质流程的共同平衡方法"干基"上是在不同的研究之间的比较有用的大多数情况下，因为它消除了原料的不同水分含量的影响。然而，应该指出的是，从实验与特定湿润原料这些计算值不一定反光贴在实验前t时的行为和这个特定原料的产率，如果它完全被物理干燥装置。众所周知，水分影响热解23的产率分布，这应该评价和比较"干"余额时牢记。 此外，由于矿物最终主要在炭和类似掩盖的结果向初始水分含量"干基"上质量平衡是不合适的高灰分含量的原料。同样地水，矿物质影响实际的热解反应的网络，因为它们促进二次热解反应，从而导致更高的炭和低生物油的产量。这样的效果，如果余额为灰分纠正只能在科学的基础上进行评估。实现这一目标的方法之一是通过建立碳平衡。从图2和图4的比较，可以看出，增加的固体一愕相比，芒草麦秸裂解后观察到的LD不仅由于该回收用焦炭的无机材料，也由于过程期间形成的有机固体的增加部分。 元素碳平衡的另一个优点是，以显示生物碳， 也就是说 ，其在所回收的产品级分分配的命运。这是更复杂的转换链的评价重要- 例如 ，热解，气化和合成如在这里介绍的情况下-由于生物碳应当尽可能有效地使用。一个在将来基于生物经济的生物质的最重要的作用是提供一个宽范围的商品提供生物碳，从而从化石资源置换碳。 一种用于在双螺杆混合反应器快速热解所提出的协议可在一些调整不同的尺度来实现。 ŧ他提出了一个单位的情况下，以10kg小时的加料容量-1已被证明是操作的复杂性和用于处理行为有意义的结果之间的可行的折衷。它既可用于不同类型的生物量和工艺条件优化筛选应用。测试具体的生物质原料是至关重要的，因为如果粗的固体残渣在热载波周期积累一定的原料特征可能导致不利的工艺操作。这样的积累没有为在结果部分提供的生物量观察到的，但它已被观察到非常硬生物材料具有大粒径（&gt; 1毫米）这限制了呈现过程的适用性。这个问题可以用一个不同的设计的热载体循环， 例如 ，通过具有同时部分燃烧热载体的气动运输减少。

作为一种替代化石碳源的利用生物质正在成为社会减少活动对地球气候的影响越来越重要。存在其他可再生能源，例如风能和太阳能，生物质能却代表了唯一可再生的碳源日期。因此，最有效地利用生物质是在生产化学品和专门液体燃料。残余的生物质应当以减少饲料，食品和化学品/燃料生产之间的竞争中。这些残基通常具有低堆积密度，从而呈现用于工业规模应用的后勤挑战。 为了应对这些挑战，bioliq概念一直站在技术1的卡尔斯鲁厄理工学院的发展。它具有一个分散的第一步废弃生物质转化成能量密集的中间（生物泥浆），随后转换在中央气化单元合成气体和最终合成所需产物（S）。气化和合成单元可以设计上所要求的工业规模在同一部位以实现商业运营。这个概念允许对不同的产品，从插入式燃料专门的燃料添加剂和散装化学品2-5。本文提出的第一个步骤，其中快速热解是用于废弃生物质转化为中间生物泥浆。在惰性气氛中快速热解的特征在于生物质的快速加热至通常450-500℃，&lt;2秒6的产生热解蒸汽的停留时间的反应温度。最常见的是，流化床反应器用于执行快速热解，但也存在特别适于以优化反应条件7不同的反应器设计。在下面给出的工作已经用双螺杆混炼反应器中进行。它提出了一个强大的技术，早已蜂Ñ ​​施加在工业规模上用于煤的热解和中试规模的油砂8。 双螺杆混合反应器的目的是与固体，预热热载体以混合固体生物质进料。混合需要是，以达到加热速率所必需的快速热解的条件下将所述生物量足以彻底。此外，无论是生物量和热载体颗粒的尺寸必须小，以实现高的传热系数和短粒子加热周期。在催化研究和技术工艺的卡尔斯鲁厄理工学院（KIT），以10kg小时的生物质输入容量的工艺开发装置协会（IKFT）-1已运作超过十年。它使用钢球作为热载体，其与一个斗式提升机的内部再循环，并与一个电加热系统重新加热。其主要目的是太子港的调查这是适合于在气化器中使用的产品和其适宜的为广泛原料9-11的验证NIQUE产物回收技术。一个更大的试验厂始建于平行于这些研究按500千克-1，这已经运营了五年的生物质输入量。它利用砂作为热载体，它是由一个热提升用气体气动再循环和通过夹带炭颗粒1,12-的部分燃烧另外加热。实验方法的以下描述是基于后其产品回收部分进行了翻新，以更好地类似于试工厂设计13的更小的工艺显影单元上。该实验装置的流程图如图1所示。 需要注意的快速热解生物油（FPBO）产品要求在气化炉的使用是很重要的是那些传统FPB制定了不同的0，这是通常用于直接燃料应用14。最重要的是，FPBO的固体含量不必须是非常低的。事实上，这是可取的，以便增加碳的可用于气化和滴在燃料后续合成的量来混合来自转换处理得到的焦炭产生的FPBO。这些事实对于理解在这里介绍的实验装置和快速热解实验别处公布的设计的区别非常重要。另一个重要的区别是，所研究的生物质转化概念为农业残余物如小麦秆是专门设计的。通常情况下，这种原料含有灰分的很大一部分。灰分是众所周知的显著影响快速热解产物分布。它导致有机缩合物（OC），并增加了固态和气态产物10,15,16的降低。这些事实都占为在这里介绍的实验装置和整个过程链的设计。大部分工业设施与低灰分含量木材经营，只是燃烧固体内部。这导致外用额外生产的热量。当使用的原料具有高灰分，char是副产品一个显著应有效地13使用。

<table border="0" cellpadding="0" cellspacing="0" width="1014">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Wheat straw</td>
			<td style="width:284px;">D&#246;rrmann Kraichtal-M&#252;nzesheim</td>
			<td style="width:141px;">n/a</td>
			<td style="width:357px;">Triticum aestivum L.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Scrap wood</td>
			<td style="width:284px;">Rettenmeier Holding AG</td>
			<td style="width:141px;">n/a</td>
			<td>According to class A2 of the German scrap wood decree (AltholzV &#167;2): glued, coated, painted, or otherwise treated scrap wood without organic halogen compounds and wood preservatives</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Miscanthus</td>
			<td style="width:284px;">Hotel-Heizungsbau Kraichgau-Odenwald</td>
			<td style="width:141px;">n/a</td>
			<td>Miscanthus xGiganteus</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Ethylene glycol</td>
			<td style="width:284px;">H&#228;ffner GmbH &#38; Co KG</td>
			<td style="width:141px;">1042090220600</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Ethanol</td>
			<td style="width:284px;">H&#228;ffner GmbH &#38; Co KG</td>
			<td style="width:141px;">1026800150600</td>
			<td>Grade 99.9 %</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Nitrogen</td>
			<td style="width:284px;">KIT</td>
			<td style="width:141px;">n/a</td>
			<td>Supplied by internal nitrogen pressure system.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Pyrolysis test rig</td>
			<td style="width:284px;">self-built</td>
			<td style="width:141px;">n/a</td>
			<td>Flow scheme is illustrated in manuscript.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Name</td>
			<td style="width:284px;">Company</td>
			<td style="width:141px;">Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Analyses:</td>
			<td style="width:284px;"></td>
			<td style="width:141px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Gas chromatograph Daniel 700</td>
			<td style="width:284px;">Emerson Process Management</td>
			<td style="width:141px;">n/a</td>
			<td>Designed for this specific application by Emerson; two 20 % SF 96 columns, two HAYESEP N columns, and one MS-5A washed column; carrier gas is helium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Helium</td>
			<td style="width:284px;">Air Liquide</td>
			<td style="width:141px;">P0252L50R2A001</td>
			<td>Grade 6.0</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:232px;">Gas mixture for calibration</td>
			<td style="width:284px;">basi Sch&#246;berl GmbH &#38; Co. KG</td>
			<td style="width:141px;">FG 10002</td>
			<td>Specified gas composition: 5 % Ne, 2 % O2, 20 % CO, 30 % CO2, 5 % CH4, 5 % H2, 2 % C2H6, 0.5 % C3H8, 0.5 % C4H10, 0.5 % C5H12, remainder N2.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Neon</td>
			<td style="width:284px;">Air Liquide</td>
			<td style="width:141px;">P0890S10R2A001</td>
			<td>Grade 4.0; used as fixed reference gas flow; not necessarily required and is only given as an example for quantifying the pyrolysis gas flow.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Elementaranalysator CHN628</td>
			<td style="width:284px;">Leco Instrumente GmbH</td>
			<td style="width:141px;">622-000-000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">TGA701</td>
			<td style="width:284px;">Leco Instrumente GmbH</td>
			<td style="width:141px;">n/a</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">DIMATOC 2000</td>
			<td style="width:284px;">Dimatec</td>
			<td style="width:141px;">n/a</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Hydranal methanol dry</td>
			<td style="width:284px;">Sigma Aldrich</td>
			<td style="width:141px;">34741</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Hydranal composite V</td>
			<td style="width:284px;">Sigma Aldrich</td>
			<td style="width:141px;">34805</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">841 Titrando</td>
			<td style="width:284px;">Deutsche Metrohm GmbH &#38; Co. KG</td>
			<td style="width:141px;">2.841.0010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">774 Oven Sample Processor</td>
			<td style="width:284px;">Deutsche Metrohm GmbH &#38; Co. KG</td>
			<td style="width:141px;">2.774.0010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">800 Dosino</td>
			<td style="width:284px;">Deutsche Metrohm GmbH &#38; Co. KG</td>
			<td style="width:141px;">2.800.0010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">801 Stirrer</td>
			<td style="width:284px;">Deutsche Metrohm GmbH &#38; Co. KG</td>
			<td style="width:141px;">2.801.0010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Methanol</td>
			<td style="width:284px;">Carl Roth GmbH &#38; Co KG</td>
			<td style="width:141px;">83884</td>
			<td>99% for synthesis</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:232px;">Whatman cellulose filter grade 42</td>
			<td style="width:284px;">Sigma Aldrich</td>
			<td style="width:141px;">WHA1442090</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Methanol-D4</td>
			<td style="width:284px;">Sigma Aldrich</td>
			<td align="right" style="width:141px;">151947</td>
			<td></td>
		</tr>
		<tr height="46">
			<td height="46" style="height:46px;width:232px;">3-(Trimethylsilyl)propionic-2,2,3,3-d4 acid sodium salt</td>
			<td style="width:284px;">Sigma Aldrich</td>
			<td align="right" style="width:141px;">269913</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">BZH&#160;250&#160;MHz</td>
			<td style="width:284px;">Bruker</td>
			<td style="width:141px;">n/a</td>
			<td></td>
		</tr>
	</tbody>
</table>

fast pyrolysis of biomass residues in a twin-screw mixing reactor

快速热解在商业植物被越来越多地应用于世界各地。他们对木质生物质，这对快速热解转化良好的性能完全运行。为了增加粮食生产和充满活力的和/或材料的使用生物质的协同效应，最好是利用从农业生产， 如秸秆残留物。所提出的方法适合于在工业规模上变换这样的材料。的主要特征都和从多个生物物质残基的转化质量平衡的一个实例。富含有机物的和富含水酮 - 转换之后，分馏冷凝以便检索两个凝聚施加。这种设计防止了生产快速热解生物油的呈现相分离。两相生物油可以预料，是因为秸秆生物质的典型的高灰分含量，促进在生产反应的水转换。 这两种分级浓缩，高灰分生物质的利用要求建立余额谨慎的做法。并非所有类型的余额都是有意义的，相当于从文献其他结果。不同平衡方法等，并进行了讨论，可以从它们中得到的信息。

各类生物质在IKFT / KIT当前设置成功裂解热解装置。例如，三个不同的原料（小麦秸秆，芒草，和废材）进行比较热解后关于以下描述的方法及其属性和产量。不同种类的平衡的方法中示出，并且对于其适用性朝富灰的原料进行讨论。要注意的是，根据每个级分的聚集状态的余额已计算和总结它是重要的。生物油在第一冷凝器中回收仍含有固体，其没有被旋风分离器除去。这些在天平单独标记。用于比较和统计评估，生物油的固体含量加入到从旋风分离器中回收的炭分数。 在一个"为获得"的基础上，固体的产量， 即 char经由生物油气旋和焦炭本回收的范围是从14-25％（重量）的调查原料。在两个冷凝器中回收总冷凝产率范围为53-66％（重量），而气体产率相对相似（约20％）对所有3生物量（ 见图2）。这些&#39;作为接收&#39;值给出对产品级分的实际量的实际信息，以在这种快速热解装置可以预期的。 然而，在文献中总液体有机产率最常报道在干燥的基础上， 即不包括在冷凝和进料在反应的湿气和水。这种平衡的优点在于初始生物质的存在的水分不影响结果的事实。这种水分含量会人为地增加了产量凝在"作为收到的"Balance。为了比较起见， 图3示出有机油收率和反应水在干燥的基础上。在这项研究中，有机产油量增加（35 - 46 - 50重量％）与降低灰分含量的原料小麦秸秆（9.2 - 1.5％重量- 2.7） -芒-废柴（ 见表1）。这与其它研究10,15,16观测线。的反应的水产量在一个比较窄的范围内，从12-14％（重量）。 以干基计质量平衡仍然直接受原料的灰分含量。包含在生物量材料的矿物质会人为地增加固体的产率在两个"作为接收的"和"干"余额。因此，元素碳平衡测定，因为它们似乎更适用于评估生物量的热化学转化反应的差异（参见图4 </st荣&gt;）。变得明显，碳的较大部分中的生物油（按重量计44-54％）和以固体形式如热解炭的24-32％仅仅一个质量分数被回收。约16-19％（重量）的碳的不以固体或液体形式回收和叶植物与热分解气体。在生产厂中，该气体将被再循环用于内燃设备的能量回收的原因。只有约3-4％的碳的质量分数在水冷凝物，其具有按重量计约80％的水含量被回收。这证实这里提出的分级缩合设置的有效性。 <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td></td><td>麦草</td><td>芒草</td><td>废柴</td></tr><tr><td>水，AR </td><td> 9.6 </td><td> 10.1 </td><td> 15.2 </td></tr><tr><td>灰，D </td><td> 9.2 </td><td> 2.7 </td><td> 1.5 </td></tr><tr><td>碳，D </td><td> 46.​​1 </td><td> 48.6 </td><td> 49.8 </td></tr><tr><td>氢，C </td><td> 5.7 </td><td> 5.9 </td><td> 6.1 </td></tr><tr><td>氮，D </td><td> &lt;0.5 </td><td> &lt;0.5 </td><td> &lt;0.5 </td></tr><tr><td colspan="4"> AR:按收到，D:干基21 </td></tr></tbody></table> 所使用的不同的原料的表1中选定的特性。所有的数值表示质量分数（％）。 <img alt="图1" src="/files/ftp_upload/54395/54395fig1.jpg" /> 图1. 实验装置。1）生物量的存储的流程图 。 2）锁斗系统。 3）生物质剂量。 4）双螺杆混合反应器。 5）斗式提升。 6）加热器热载体。 7）旋风固体去除。 8）字符的存储。 9）喷淬火。 10）生物油储存罐。 11）均质和泵。 12）换热器用于再循环冷凝物的冷却。 13）电除尘器。 14）水性冷凝储罐。 15）泵的循环水冷凝水。 16）换热器的冷凝循环冷却。 17）为冷凝器冷凝水。 18）风扇去除燃气/蒸汽。 <a href="https://www.jove.com/files/ftp_upload/54395/54395fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/54395/54395fig2.jpg" /> 图2 的实验的质量平衡。天平上报告的原料和产品的"所接收"21的基础。所有值表示为质量份。三种不同类型的生物质已被用于所有实验至少重复三次13进行。在生物的固体含量油是用于说明目的单独报告。误差棒表示与一类原料的实验标准差。 <a href="https://www.jove.com/files/ftp_upload/54395/54395fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/54395/54395fig3.jpg" /> 图3 总的有机油产率和反应的水。所有值都以干21进料的基础上，并表示为质量份。冷凝物的固体含量已被排除，从有机油收率13。误差棒表示与一类原料的实验标准差。 <a href="https://www.jove.com/files/ftp_upload/54395/54395fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 图4. 碳余额。所有值报告为生物量的碳输入的质量分数。三种不同类型的生物质已被用于所有实验至少重复三次13进行。在生物油中的固体含量为说明目的单独报告。误差棒表示与一类原料的实验标准差。 <a href="https://www.jove.com/files/ftp_upload/54395/54395fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Fast Pyrolysis of Biomass Residues in a Twin-screw Mixing Reactor at JoVE.com

Fast Pyrolysis of Biomass Residues in a Twin-screw Mixing Reactor

A procedure for thermochemical conversion of biomass residues is presented that aims at maximizing the yield of liquid products (fast pyrolysis). It is based on a technology proven on an industrial scale and especially suitable for treating a straw type of biomass.

在双螺杆混炼反应器生物残渣快速热解

Fast pyrolysis is being increasingly applied in commercial plants worldwide. They run exclusively on woody biomass, which has favorable properties for ...

fast-pyrolysis-of-biomass-residues-in-a-twin-screw-mixing-reactor

Research

JoVE Journal

Bioengineering

27.5K 次观看.  Karlsruhe Institute of Technology (KIT). A procedure for thermochemical conversion of biomass residues is presented that aims at maximizing the yield of liquid products (fast pyrolysis). It is based on a technology proven on an industrial scale and especially suitable for treating a straw type of biomass.

视频: Fast Pyrolysis of Biomass Residues in a Twin-screw Mixing Reactor

Creating Two-Dimensional Patterned Substrates for Protein and Cell Confinement

Microcontact printing provides a rapid, highly reproducible method for the creation of well-defined patterned substrates.1 While microcontact printing can be employed to directly print a large number of molecules, including proteins,2 DNA,3 and silanes,4 the formation of self-assembled monolayers (SAMs) from long chain alkane thiols on gold provides a simple way to confine proteins and cells to specific patterns containing adhesive and resistant regions. This confinement can be used to control cell morphology and is useful for examining a variety of questions in protein and cell biology. Here, we describe a general method for the creation of well-defined protein patterns for cellular studies.5 This process involves three steps: the production of a patterned master using photolithography, the creation of a PDMS stamp, and microcontact printing of a gold-coated substrate. Once patterned, these cell culture substrates are capable of confining proteins and/or cells (primary cells or cell lines) to the pattern.
The use of self-assembled monolayer chemistry allows for precise control over the patterned protein/cell adhesive regions and non-adhesive regions; this cannot be achieved using direct protein stamping. Hexadecanethiol, the long chain alkane thiol used in the microcontact printing step, produces a hydrophobic surface that readily adsorbs protein from solution. The glycol-terminated thiol, used for backfilling the non-printed regions of the substrate, creates a monolayer that is resistant to protein adsorption and therefore cell growth.6 These thiol monomers produce highly structured monolayers that precisely define regions of the substrate that can support protein adsorption and cell growth. As a result, these substrates are useful for a wide variety of applications from the study of intercellular behavior7 to the creation of microelectronics.8
While other types of monolayer chemistry have been used for cell culture studies, including work from our group using trichlorosilanes to create patterns directly on glass substrates,9 patterned monolayers formed from alkane thiols on gold are straight-forward to prepare. Moreover, the monomers used for monolayer preparation are commercially available, stable, and do not require storage or handling under inert atmosphere. Patterned substrates prepared from alkane thiols can also be recycled and reused several times, maintaining cell confinement.10

Self-assembled monolayers (SAMs) formed from long chain alkane thiols on gold provide well-defined substrates for the formation of protein patterns and cell confinement. Microcontact printing of hexadecanethiol using a polydimethylsiloxane (PDMS) stamp followed by backfilling with a glycol-terminated alkane thiol monomer produces a pattern where protein and cells adsorb only to the stamped hexadecanethiol region.

蛋白质和细胞坐月子创建二维图案衬底

Microcontact printing provides a rapid, highly reproducible method for the creation of well-defined patterned substrates.1  While microcontact printing ...

科研

生物工程

GENPLAT: an Automated Platform for Biomass Enzyme Discovery and Cocktail Optimization

The high cost of enzymes for biomass deconstruction is a major impediment to the economic conversion of lignocellulosic feedstocks to liquid transportation fuels such as ethanol. We have developed an integrated high throughput platform, called GENPLAT, for the discovery and development of novel enzymes and enzyme cocktails for the release of sugars from diverse pretreatment/biomass combinations. GENPLAT comprises four elements: individual pure enzymes, statistical design of experiments, robotic pipeting of biomass slurries and enzymes, and automated colorimeteric determination of released Glc and Xyl. Individual enzymes are produced by expression in Pichia pastoris or Trichoderma reesei, or by chromatographic purification from commercial cocktails or from extracts of novel microorganisms. Simplex lattice (fractional factorial) mixture models are designed using commercial Design of Experiment statistical software. Enzyme mixtures of high complexity are constructed using robotic pipeting into a 96-well format. The measurement of released Glc and Xyl is automated using enzyme-linked colorimetric assays. Optimized enzyme mixtures containing as many as 16 components have been tested on a variety of feedstock and pretreatment combinations.
GENPLAT is adaptable to mixtures of pure enzymes, mixtures of commercial products (e.g., Accellerase 1000 and Novozyme 188), extracts of novel microbes, or combinations thereof. To make and test mixtures of &tilde;10 pure enzymes requires less than 100 &mu;g of each protein and fewer than 100 total reactions, when operated at a final total loading of 15 mg protein/g glucan. We use enzymes from several sources. Enzymes can be purified from natural sources such as fungal cultures (e.g., Aspergillus niger, Cochliobolus carbonum, and Galerina marginata), or they can be made by expression of the encoding genes (obtained from the increasing number of microbial genome sequences) in hosts such as E. coli, Pichia pastoris, or a filamentous fungus such as T. reesei. Proteins can also be purified from commercial enzyme cocktails (e.g., Multifect Xylanase, Novozyme 188). An increasing number of pure enzymes, including glycosyl hydrolases, cell wall-active esterases, proteases, and lyases, are available from commercial sources, e.g., Megazyme, Inc. (www.megazyme.com), NZYTech (www.nzytech.com), and PROZOMIX (www.prozomix.com).
Design-Expert software (Stat-Ease, Inc.) is used to create simplex-lattice designs and to analyze responses (in this case, Glc and Xyl release). Mixtures contain 4-20 components, which can vary in proportion between 0 and 100%. Assay points typically include the extreme vertices with a sufficient number of intervening points to generate a valid model. In the terminology of experimental design, most of our studies are "mixture" experiments, meaning that the sum of all components adds to a total fixed protein loading (expressed as mg/g glucan). The number of mixtures in the simplex-lattice depends on both the number of components in the mixture and the degree of polynomial (quadratic or cubic). For example, a 6-component experiment will entail 63 separate reactions with an augmented special cubic model, which can detect three-way interactions, whereas only 23 individual reactions are necessary with an augmented quadratic model. For mixtures containing more than eight components, a quadratic experimental design is more practical, and in our experience such models are usually statistically valid.
All enzyme loadings are expressed as a percentage of the final total loading (which for our experiments is typically 15 mg protein/g glucan). For "core" enzymes, the lower percentage limit is set to 5%. This limit was derived from our experience in which yields of Glc and/or Xyl were very low if any core enzyme was present at 0%. Poor models result from too many samples showing very low Glc or Xyl yields. Setting a lower limit in turn determines an upper limit. That is, for a six-component experiment, if the lower limit for each single component is set to 5%, then the upper limit of each single component will be 75%. The lower limits of all other enzymes considered as "accessory" are set to 0%. "Core" and "accessory" are somewhat arbitrary designations and will differ depending on the substrate, but in our studies the core enzymes for release of Glc from corn stover comprise the following enzymes from T. reesei: CBH1 (also known as Cel7A), CBH2 (Cel6A), EG1(Cel7B), BG (&beta;-glucosidase), EX3 (endo-&beta;1,4-xylanase, GH10), and BX (&beta;-xylosidase).
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GENPLAT (GLBRC Enzyme Platform) is an automated platform for discovery and optimization of enzyme cocktails for biomass degradation. It can be adapted to multiple feedstocks and mixtures of enzymes containing multiple components.

GENPLAT:生物质酶的发现和鸡尾酒优化的自动化平台

The high cost of enzymes for biomass deconstruction is a major impediment to the economic conversion of lignocellulosic feedstocks to liquid ...

Adaptation of a Haptic Robot in a 3T fMRI

Functional magnetic resonance imaging (fMRI) provides excellent functional brain imaging via the BOLD signal 1 with advantages including non-ionizing radiation, millimeter spatial accuracy of anatomical and functional data 2, and nearly real-time analyses 3. Haptic robots provide precise measurement and control of position and force of a cursor in a reasonably confined space. Here we combine these two technologies to allow precision experiments involving motor control with haptic/tactile environment interaction such as reaching or grasping. The basic idea is to attach an 8 foot end effecter supported in the center to the robot 4 allowing the subject to use the robot, but shielding it and keeping it out of the most extreme part of the magnetic field from the fMRI machine (Figure 1).
The Phantom Premium 3.0, 6DoF, high-force robot (SensAble Technologies, Inc.) is an excellent choice for providing force-feedback in virtual reality experiments 5, 6, but it is inherently non-MR safe, introduces significant noise to the sensitive fMRI equipment, and its electric motors may be affected by the fMRI's strongly varying magnetic field. We have constructed a table and shielding system that allows the robot to be safely introduced into the fMRI environment and limits both the degradation of the fMRI signal by the electrically noisy motors and the degradation of the electric motor performance by the strongly varying magnetic field of the fMRI. With the shield, the signal to noise ratio (SNR: mean signal/noise standard deviation) of the fMRI goes from a baseline of &tilde;380 to &tilde;330, and &tilde;250 without the shielding. The remaining noise appears to be uncorrelated and does not add artifacts to the fMRI of a test sphere (Figure 2). The long, stiff handle allows placement of the robot out of range of the most strongly varying parts of the magnetic field so there is no significant effect of the fMRI on the robot. The effect of the handle on the robot's kinematics is minimal since it is lightweight (&tilde;2.6 lbs) but extremely stiff 3/4" graphite and well balanced on the 3DoF joint in the middle. The end result is an fMRI compatible, haptic system with about 1 cubic foot of working space, and, when combined with virtual reality, it allows for a new set of experiments to be performed in the fMRI environment including naturalistic reaching, passive displacement of the limb and haptic perception, adaptation learning in varying force fields, or texture identification 5, 6.

The adaptation and use of a haptic robot in a 3T fMRI is described.

触觉机器人适应在3T功能磁共振成像

Functional magnetic resonance imaging (fMRI) provides excellent functional brain imaging via the BOLD signal 1 with advantages including non-ionizing ...

Cellular Lipid Extraction for Targeted Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry Analysis

The metabolism of fatty acids, such as arachidonic acid (AA) and linoleic acid (LA), results in the formation of oxidized bioactive lipids, including numerous stereoisomers1,2. These metabolites can be formed from free or esterified fatty acids. Many of these oxidized metabolites have biological activity and have been implicated in various diseases including cardiovascular and neurodegenerative diseases, asthma, and cancer3-7. Oxidized bioactive lipids can be formed enzymatically or by reactive oxygen species (ROS). Enzymes that metabolize fatty acids include cyclooxygenase (COX), lipoxygenase (LO), and cytochromes P450 (CYPs)1,8. Enzymatic metabolism results in enantioselective formation whereas ROS oxidation results in the racemic formation of products.
While this protocol focuses primarily on the analysis of AA- and some LA-derived bioactive metabolites; it could be easily applied to metabolites of other fatty acids. Bioactive lipids are extracted from cell lysate or media using liquid-liquid (l-l) extraction. At the beginning of the l-l extraction process, stable isotope internal standards are added to account for errors during sample preparation. Stable isotope dilution (SID) also accounts for any differences, such as ion suppression, that metabolites may experience during the mass spectrometry (MS) analysis9. After the extraction, derivatization with an electron capture (EC) reagent, pentafluorylbenzyl bromide (PFB) is employed to increase detection sensitivity10,11. Multiple reaction monitoring (MRM) is used to increase the selectivity of the MS analysis. Before MS analysis, lipids are separated using chiral normal phase high performance liquid chromatography (HPLC). The HPLC conditions are optimized to separate the enantiomers and various stereoisomers of the monitored lipids12. This specific LC-MS method monitors prostaglandins (PGs), isoprostanes (isoPs), hydroxyeicosatetraenoic acids (HETEs), hydroxyoctadecadienoic acids (HODEs), oxoeicosatetraenoic acids (oxoETEs) and oxooctadecadienoic acids (oxoODEs); however, the HPLC and MS parameters can be optimized to include any fatty acid metabolites13.
Most of the currently available bioanalytical methods do not take into account the separate quantification of enantiomers. This is extremely important when trying to deduce whether or not the metabolites were formed enzymatically or by ROS. Additionally, the ratios of the enantiomers may provide evidence for a specific enzymatic pathway of formation. The use of SID allows for accurate quantification of metabolites and accounts for any sample loss during preparation as well as the differences experienced during ionization. Using the PFB electron capture reagent increases the sensitivity of detection by two orders of magnitude over conventional APCI methods. Overall, this method, SID-LC-EC-atmospheric pressure chemical ionization APCI-MRM/MS, is one of the most sensitive, selective, and accurate methods of quantification for bioactive lipids.

This protocol will demonstrate the extraction and analysis of free and esterified bioactive fatty acids from cells. Fatty acids are accurately quantified using stable isotope dilution, chiral liquid chromatography, electron capture atmospheric chemical ionization multiple reaction monitoring mass spectrometry (SID-LC-ECAPCI-MRM/MS).

有针对性的稳定同位素稀释液相色谱 - 质谱法分析细胞脂质提取

The metabolism of fatty acids, such as arachidonic acid (AA) and linoleic acid (LA), results in the formation of oxidized bioactive lipids, including ...

Quantification of Global Diastolic Function by Kinematic Modeling-based Analysis of Transmitral Flow via the Parametrized Diastolic Filling Formalism

Quantitative cardiac function assessment remains a challenge for physiologists and clinicians.&#160;Although historically invasive methods have comprised the only means available, the development of noninvasive imaging modalities (echocardiography, MRI, CT) having high temporal and spatial resolution provide a new window for quantitative diastolic function assessment. Echocardiography is the agreed upon standard for diastolic function assessment, but indexes in current clinical use merely utilize selected features of chamber dimension (M-mode) or blood/tissue motion (Doppler) waveforms without incorporating the physiologic causal determinants of the motion itself.&#160;The recognition that all left ventricles (LV) initiate filling by serving as mechanical suction pumps allows global diastolic function to be assessed based on laws of motion that apply to all chambers. What differentiates one heart from another are the parameters of the equation of motion that governs filling. Accordingly, development of the Parametrized Diastolic Filling (PDF) formalism&#160;has shown that the entire range of clinically observed early transmitral flow (Doppler E-wave) patterns are extremely well fit by the laws of damped oscillatory motion.&#160;This permits analysis of individual E-waves in accordance with a causal mechanism (recoil-initiated suction) that yields three (numerically) unique lumped parameters whose physiologic analogues are chamber stiffness (k), viscoelasticity/relaxation (c), and load (xo).&#160;The recording of transmitral flow (Doppler E-waves) is standard practice in clinical cardiology and, therefore, the echocardiographic recording method is only briefly reviewed. Our focus is on determination of the PDF parameters from routinely recorded E-wave data.&#160;As the highlighted results indicate, once the PDF parameters have been obtained from a suitable number of load varying E-waves, the investigator is free to use the parameters or construct indexes from the parameters (such as stored energy 1/2kxo2, maximum A-V pressure gradient kxo, load independent index of diastolic function, etc.) and select the aspect of physiology or pathophysiology to be quantified.

Accurate, causality-based quantification of global diastolic function has been achieved by kinematic modeling-based analysis of transmitral flow via the Parametrized Diastolic Filling (PDF) formalism. PDF generates unique stiffness, relaxation,&#160;and load parameters and elucidates &#39;new&#39; physiology while providing sensitive and specific indexes of dysfunction.

全球舒张功能通过静脉流的通过参数化舒张形式主义运动学建模为基础的定量分析

Quantitative cardiac function assessment remains a challenge for physiologists and clinicians.&#160;Although historically invasive methods have comprised ...

From Fast Fluorescence Imaging to Molecular Diffusion Law on Live Cell Membranes in a Commercial Microscope

It has become increasingly evident that the spatial distribution and the motion of membrane components like lipids and proteins are key factors in the regulation of many cellular functions. However, due to the fast dynamics and the tiny structures involved, a very high spatio-temporal resolution is required to catch the real behavior of molecules. Here we present the experimental protocol for studying the dynamics of fluorescently-labeled plasma-membrane proteins and lipids in live cells with high spatiotemporal resolution. Notably, this approach doesn&#8217;t need to track each molecule, but it calculates population behavior using all molecules in a given region of the membrane. The starting point is a fast imaging of a given region on the membrane. Afterwards, a complete spatio-temporal autocorrelation function is calculated correlating acquired images at increasing time delays, for example each 2, 3, n repetitions. It is possible to demonstrate that the width of the peak of the spatial autocorrelation function increases at increasing time delay as a function of particle movement due to diffusion. Therefore, fitting of the series of autocorrelation functions enables to extract the actual protein mean square displacement from imaging (iMSD), here presented in the form of apparent diffusivity vs average displacement. This yields a quantitative view of the average dynamics of single molecules with nanometer accuracy. By using a GFP-tagged variant of the Transferrin Receptor (TfR) and an ATTO488 labeled 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphoethanolamine (PPE) it is possible to observe the spatiotemporal regulation of protein and lipid diffusion on &#181;m-sized membrane regions in the micro-to-milli-second time range.

Spatial distribution and temporal dynamics of plasma membrane proteins and lipids is a hot topic in biology. Here this issue is addressed by a spatio-temporal image fluctuation analysis that provides conceptually the same physical quantities of single particle tracking, but it uses small molecular labels and standard microscopy setups.

从快速荧光成像在分子扩散法对活细胞的膜在商业显微镜

It has become increasingly evident that the spatial distribution and the motion of membrane components like lipids and proteins are key factors in the ...

Analyzing Mixing Inhomogeneity in a Microfluidic Device by Microscale Schlieren Technique

In this paper, we introduce the use of microscale schlieren technique to measure mixing inhomogeneity in a microfluidic device. The microscale schlieren system is constructed from a Hoffman modulation contrast microscope, which provides easy access to the rear focal plane of the objective lens, by removing the slit plate and replacing the modulator with a knife-edge. The working principle of microscale schlieren technique relies on detecting light deflection caused by variation of refractive index1-3. The deflected light either escapes or is obstructed by the knife-edge to produce a bright or a dark band, respectively. If the refractive index of the mixture varies linearly with its composition, the local change in light intensity in the image plane is proportional to the concentration gradient normal to the optical axis. The micro-schlieren image gives a two-dimensional projection of the disturbed light produced by three-dimensional inhomogeneity.
To accomplish quantitative analysis, we describe a calibration procedure that mixes two fluids in a T-microchannel. We carry out a numerical simulation to obtain the concentration gradient in the T-microchannel that correlates closely with the corresponding micro-schlieren image. By comparison, a relationship between the grayscale readouts of the micro-schlieren image and the concentration gradients presented in a microfluidic device is established. Using this relationship, we are able to analyze the mixing inhomogeneity from associate micro-schlieren image and demonstrate the capability of microscale schlieren technique with measurements in a microfluidic oscillator4. For optically transparent fluids, microscale schlieren technique is an attractive diagnostic tool to provide instantaneous full-field information that retains the three-dimensional features of the mixing process.

Herein, we describe a procedure that employs microscale schlieren technique to measure mixing inhomogeneity in a microfluidic device. Through calibration, distribution of concentration gradient can be derived from the micro-schlieren image.

由微型条纹分析技术在微流控设备混合不均匀

In this paper, we introduce the use of microscale schlieren technique to measure mixing inhomogeneity in a microfluidic device. The microscale schlieren ...

Microwave-driven Synthesis of Iron Oxide Nanoparticles for Fast Detection of Atherosclerosis

A fast and reproducible microwave-driven protocol has been developed for the synthesis of neridronate-functionalized nanoparticles. Starting from the synthesis of hydrophobic nanoparticles, our method is based on an adaptation from thermal decomposition method to microwave driven synthesis. The new methodology produces a decrease in the reaction times in comparison with traditional procedures. Moreover, the use of the microwave technology increases the reproducibility of the reactions, something important from the point of view of clinical applications. The novelty of this iron oxide nanoparticle is the attachment of Neridronate. The use of this molecule leads a bisphosphonate moiety towards the outside of the nanoparticle that provides Ca2+ binding properties in vitro and selective accumulation in vivo in the atheroma plaque. The protocol allows the synthesis and plaque detection in about 3 hr since the initial synthesis from organic precursors. Their accumulation in the atherosclerotic area in less than 1 hr provides a contrast agent particularly suitable for clinical applications.

Microwave technology enables extremely fast synthesis of iron oxide nanoparticles for atherosclerosis plaque characterization. The use of an aminobisphosphonate in the external side of the nanoparticle provides a fast accumulation in the atherosclerotic area.

动脉粥样硬化的快速检测的氧化铁纳米颗粒微波驱动综合

A fast and reproducible microwave-driven protocol has been developed for the synthesis of neridronate-functionalized nanoparticles. Starting from the ...

Fast Enzymatic Processing of Proteins for MS Detection with a Flow-through Microreactor

The vast majority of mass spectrometry (MS)-based protein analysis methods involve an enzymatic digestion step prior to detection, typically with trypsin. This step is necessary for the generation of small molecular weight peptides, generally with MW &#60; 3,000-4,000 Da, that fall within the effective scan range of mass spectrometry instrumentation. Conventional protocols involve O/N enzymatic digestion at 37 &#186;C. Recent advances have led to the development of a variety of strategies, typically involving the use of a microreactor with immobilized enzymes or of a range of complementary physical processes that reduce the time necessary for proteolytic digestion to a few minutes (e.g., microwave or high-pressure). In this work, we describe a simple and cost-effective approach that can be implemented in any laboratory for achieving fast enzymatic digestion of a protein. The protein (or protein mixture) is adsorbed on C18-bonded reversed-phase high performance liquid chromatography (HPLC) silica particles preloaded in a capillary column, and trypsin in aqueous buffer is infused over the particles for a short period of time. To enable on-line MS detection, the tryptic peptides are eluted with a solvent system with increased organic content directly in the MS ion source. This approach avoids the use of high-priced immobilized enzyme particles and does not necessitate any aid for completing the process. Protein digestion and complete sample analysis can be accomplished in less than ~3 min and ~30 min, respectively.

A quick protocol for proteolytic digestion with an in-house built flow-through tryptic microreactor coupled to an electrospray ionization (ESI) mass spectrometer is presented. The fabrication of the microreactor, the experimental setup and the data acquisition process are described.

蛋白质的MS检测快速酶法处理与流通型微反应器

The vast majority of mass spectrometry (MS)-based protein analysis methods involve an enzymatic digestion step prior to detection, typically with trypsin. ...

Synthesis of Cationized Magnetoferritin for Ultra-fast Magnetization of Cells

Many important biomedical applications, such as cell imaging and remote manipulation, can be achieved by labeling cells with superparamagnetic iron oxide nanoparticles (SPIONs). Achieving sufficient cellular uptake of SPIONs is a challenge that has traditionally been met by exposing cells to elevated concentrations of SPIONs or by prolonging exposure times (up to 72 hr). However, these strategies are likely to mediate toxicity. Here, we present the synthesis of the protein-based SPION magnetoferritin as well as a facile surface functionalization protocol that enables rapid cell magnetization using low exposure concentrations. The SPION core of magnetoferritin consists of cobalt-doped iron oxide with an average particle diameter of 8.2 nm mineralized inside the cavity of horse spleen apo-ferritin. Chemical cationization of magnetoferritin produced a novel, highly membrane-active SPION that magnetized human mesenchymal stem cells (hMSCs) using incubation times as short as one minute and iron concentrations as lows as 0.2 mM.

A protocol for the synthesis and cationization of cobalt-doped magnetoferritin is presented, as well as a method to rapidly magnetize stem cells with cationized magnetoferritin.

对细胞的超快磁化阳离子化Magnetoferritin的合成

Many important biomedical applications, such as cell imaging and remote manipulation, can be achieved by labeling cells with superparamagnetic iron oxide ...