Name: label-free in situ imaging of lignification in plant cell walls
Uploaded: 2010-11-01T17:00:00
Description: Watch this Scientific Journal Video about Label-free in situ Imaging of Lignification in Plant Cell Walls at JoVE.com

我们感谢安德鲁卡罗尔，明亮Chaibang，Purbasha萨卡（伯克利分校能源生物科学研究所），巴赫拉姆Parvin（劳伦斯伯克利国家实验室）和Vincent L.蒋（北卡罗莱纳州立大学）进行富有成效的合作和有益的讨论。这项工作是支持的能源生物科学研究所。在分子铸造的工作是支持由美国能源部科学办公室基础能源科学办公室的，根据合同号DE - AC02 - 05CH1123。

1。样品制备<ol><li>摩水合植物样品，如杨树干木， 拟南芥干，在切片机。 </li><li>从本地组织切薄片（一般为20微米厚）。 </li><li>工厂部分转移到玻璃显微镜幻灯片。 </li><li>浸泡在D 2 O的植物部分，并盖上玻璃盖滑，这是在显微镜玻片上，密封，以防止蒸发的D 2 O现在，该厂部分成像，也可以保存供日后使用。 </li></ol> 2。样品测​​定<ol><li>应用浸油，...

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	<li>Herrera, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bonkers+about+biofuels.">Bonkers about biofuels.</a> Nat Biotechnol. 24, 755-760 (2006).</li><li>Himmel, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biomass+recalcitrance:+Engineering+plants+and+enzymes+for+biofuels+production.">Biomass recalcitrance: Engineering plants and enzymes for biofuels production.</a> Science. 315, 804-807 (2007).</li><li>Pauly, M., Keegstra, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell-wall+carbohydrates+and+their+modification+as+a+resource+for+biofuels.">Cell-wall carbohydrates and their modification as a resource for biofuels.</a> Plant J. 54, 559-568 (2008).</li><li>Pauly, M., Keegstra, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physiology+and+metabolism+'Tear+down+this+wall.">Physiology and metabolism 'Tear down this wall.</a> Curr Opin Plant Biol. 11, 233-235 (2008).</li><li>Ragauskas, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+path+forward+for+biofuels+and+biomaterials.">The path forward for biofuels and biomaterials.</a> Science. 311, 484-489 (2006).</li><li>Somerville, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biofuels.">Biofuels.</a> Curr Biol. 17, R115-R119 (2007).</li><li>Ralph, J., Brunow, G., Boerjan, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Lignins in Encyclopedia of Life Sciences. , (2007).</li><li>Chiang, V. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+rags+to+riches.">From rags to riches.</a> Nat Biotechnol. 20, 557-558 (2002).</li><li>Atalla, R. H., Agarwal, U. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Raman+microprobe+evidence+for+lignin+orientation+in+the+cell+walls+of+native+woody+tissue.">Raman microprobe evidence for lignin orientation in the cell walls of native woody tissue.</a> Science. 227, 636-638 (1985).</li><li>Atalla, R. H., Agarwal, U. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recording+Raman+spectra+from+plant+cell+walls.">Recording Raman spectra from plant cell walls.</a> J Raman Spectrosc. 17, 229-231 (1986).</li><li>Fukushima, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+syringyl+to+guaiacyl+ratio+in+lignin+biosynthesis.">Regulation of syringyl to guaiacyl ratio in lignin biosynthesis.</a> J Plant Res. 114, 499-508 (2001).</li><li>Agarwal, U. 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木质纤维素材料的层次和异构的结构和组成方面。在深入刻画的分析工具，化学灵敏度，空间分辨率，而且这些材料在本地范围内的见解是可取的。所描述的方法，让该可视化与空间分辨率，是亚微米无染色或以接近原生状态的样品标签木质纤维植物生物量的木质化，木质素和比较。它需要最少的样品制备和测量都是非破坏性和相对低廉。该方法可在评估大量的繁殖种群的木质化的有用。此外，?...

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<td>microscope slides</td>
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<td>D2O</td>
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<td>immersion oil</td>
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<td>tweezers</td>
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<td>pointed brush</td>
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<td>microtome</td>
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label-free in situ imaging of lignification in plant cell walls

满足日益增长的能源需求安全，高效，是一项紧迫的全球性挑战。因此，为生产生物燃料的研究，力求找到符合成本效益和可持续的解决方案已经成为一个局部和关键任务。将木质纤维素生物质的液体生物燃料1-6转换成为生物质的主要来源。然而，这些植物细胞壁材料的顽抗，以成本效益和高效率的降解，在生物燃料和化学品4的生产供其使用的一个主要障碍。特别是，木质素，聚苯丙heteropolymer一个复杂和不规则，成为问题的木质纤维素生物质的采后解构。例如，在生物质转化为生物燃料，它能够抑制糖化，目的是简单的糖发酵7生产的进程。用于工业用途的植物生物量的有效利用，其实在很大程度上依赖，植物细胞壁的木质化程度。去除木质素是昂贵的和限制因素和木质素，因此成为一个重要的植物育种和基因工程的目标，以改善细胞壁的转换。 评估大量的繁殖种群，允许植物细胞壁的木质化的快速准确的表征的分析工具变得越来越重要。本机组件，如木质素的分离提取过程是不可避免的破坏性，带来显着的化学和结构修改9-11。分析化学原位方法，因此木质纤维材料的成分和结构表征的宝贵工具。拉曼光谱是一种技术，依赖于无弹性或拉曼散射的单色光，如激光，在激光光子能量转变为分子振动，并提出一种内在的无标记分子的“指纹”的样本。拉曼显微镜可以承受最小的样品制备和相对低廉的非破坏性测量，给人以接近原生状态转化为化学成分和分子结构的见解。化学共焦显微拉曼光谱成像先前已用于可视化的空间分布在木材细胞墙壁12-14中的纤维素和木质素。基于这些早期的结果，我们最近通过这种方法比较野生型和木质素缺陷的转基因杨树毛果（黑三角叶）干木材 15木质化。分析木质素拉曼带在1600和1700 cm - 1处，木质素的信号强度和本地化之间的光谱区域 16,17 映射到原位。我们的方法可视化木质素含量的差异，本地化和化学成分。最近，我们展示了在拟南芥细胞壁聚合物的拉曼成像横向分辨率，亚微米 18 。在这里，这种方法是在植物细胞壁的木质素的可视化和木质化比较在不同的组织，样品或物种affording无染色或组织标签。

Watch this Scientific Journal Video about Label-free in situ Imaging of Lignification in Plant Cell Walls at JoVE.com

Label-free in situ Imaging of Lignification in Plant Cell Walls

基于共焦显微拉曼光谱方法是给予无标签的可视化在植物细胞壁的木质素和木质化比较在不同的组织，样品或物种。

无标签原位成像在植物细胞壁的木质化

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Label-free in situ Imaging of Lignification in Plant Cell Walls

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