VISUALIZE MARINE DIATOM ULTRASTRUCTURE OR EXTRACELLULAR MATRIX BY CRYO STEM

通过冷冻干可视化海洋硅藻超微结构或细胞外基质

• 批准号: 6117289
• 负责人: MICHAEL R GRETZ
• 金额: $ 1.13万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6117289
• 关键词: bioimaging /biomedical imaging; biomaterials; biomedical resource; carbohydrates; growth /development; microscopy; plants; proteins; technology /technique

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.

我们目前由海军研究办公室资助 研究藻类生物粘合剂的生产。 我们的项目 涉及生物污染硅藻 Achnanthes longipes。 上的 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 细胞外粘合剂。 为了实现这个目标我们 使用了多种技术，包括 DIC 视频延时和 荧光显微镜与传统电子显微镜相结合 显微技术。 我们还制定/实施了具体的 针对粘合剂的分子探针，包括单克隆 抗体、果胶和酶。 虽然我们感兴趣 我们对其他可用技术的研究的潜在应用 在 HAR，我们现在需要两种方法。 的粘合剂 芨芨草非常亲水并且经历显着的形态变化 暴露于标准电子显微镜脱水时发生变化 TEM 和 SEM 的程序。 为了观察散装粘合剂 如果没有这些脱水伪影，我们就不需要冷冻扫描电镜技术 可在 NITU 获取。 我们在以下方面也基本上不成功 可视化粘合剂合成的部位，以进一步 使用标准 TEM 固定来表征 inh-无细胞 技术。 我们希望尝试高压冷冻芨芨草 积极参与粘合剂合成，目标是进一步 表征所涉及的细胞内结构。 我们计划 将活组织带到 IMR 进行冷冻扫描电镜 现场观察。 我们还想利用 IMR 设施 进行高压冷冻，组织将被进一步处理 MTU 的 TEM。