VISUALIZE MARINE DIATOM ULTRASTRUCTURE OR EXTRACELLULAR MATRIX BY CRYO STEM

我们目前由海军研究办公室资助，以 对藻类生产生物粘合剂进行了研究。我们的项目 涉及生物污染的硅藻长鞭毛藻。上一次 本研究的目标是确定结构和 硅藻特定成分的生物发生机制 胞外粘合剂。为了实现这一目标，我们 利用了几种技术，包括DIC视频延时和 荧光显微镜与传统电子的耦合 显微技术。我们还开发/实施了特定的 针对包括单抗在内的粘合剂的分子探针 抗体、果胶和酶。尽管我们感兴趣的是 对我们研究其他可用技术的潜在应用 在HAR，我们现在需要两种方法。的胶粘剂 鞭毛虫是非常亲水的，经历了显著的形态 暴露于标准电子显微镜脱水时的变化 透射电子显微镜和扫描电子显微镜检查程序。为了观察散装胶粘剂 没有这些脱水文物，我们需要低温扫描技术，而不是 在Nitu有售。我们在很大程度上也没有成功 可视化粘合剂合成的位置，目标是进一步 应用标准的透射电子显微镜固定技术对股骨头脱细胞进行表征 技巧。我们希望尝试对阿奇纳提斯进行高压冷冻 积极参与胶粘剂合成，目标是进一步 描述所涉及的细胞内结构。我们计划 将活组织带到IMR，打算做冷冻扫描 现场观察。我们还希望利用IMR设施来 做高压冷冻，组织就会进一步加工 对于MTU的瞬变电磁来说。

We are currently funded by the Office of Naval Research to investigate the production of bioadhesives by algae. Our project involves the biofouling diatom Achnanthes longipes. On of the objectives in this research is to determine the structure and mechanism of biogenesis of the specific components of diatom extracellular adhesives. In order to accomplish this ob ective we have utilized several techniques including DIC video time-lapse and fluorescence microscopy coupled with traditional electron microscopical techniques. We have also developed/implemented specific molecular probes directed against the adhesives including monoclonal antibodies, pectins and enzymes. Although we are interested in potential applications to our research of other techniques available at the HAR, we have need for two methods now. The adhesives of Achnanthes are very hydrophilic and undergo significant morphological change when exposed to standard electron microscopical dehydration procedures for TEM and SEM. In order to observe the bulk adhesive without these dehydration artifacts we require cryo-SEM technology not available at NITU. We also have been largely unsuccessful at visualizing the site of adhesive synthesis with the goal of further characterizing the inh-acellularly using standard TEM fixation techniques. We wish to attempt high-pressure freezing of Achnanthes actively involved in adhesive synthesis with the goal of further characterizing the intracellular structures involved. We plan to bring live tissues to the IMR with the intent to do the cryo-SEM observations on site. We would also like to utilize IMR facilities to do high pressure freezing and the tissue would be further processed for TEM at MTU.