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: 6278484
• 负责人: MICHAEL R GRETZ
• 金额: $ 0.4万
• 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: 1998-07-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6278484
• 关键词: 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， 研究藻类生产生物粘合剂。 我们的项目 涉及生物污染硅藻长柄曲壳藻 银行或公司的 本研究的目的是确定结构和 硅藻特殊组分的生物成因机制 细胞外粘合剂 为了实现这一目标，我们 已经利用了几种技术，包括DIC视频延时， 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。 尽管我们对 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。 的粘合剂 曲壳藻是非常亲水的， 暴露于标准电子显微镜脱水时的变化 TEM和SEM的方法。 为了观察散装粘合剂 如果没有这些脱水人工制品，我们需要低温SEM技术， 在NITU。 我们也在很大程度上不成功， 可视化粘合剂合成的位点，目的是进一步 使用标准TEM固定表征inh-非细胞 技术. 我们希望尝试高压冷冻曲壳藻 积极参与粘合剂合成， 表征所涉及的细胞内结构。 我们计划 将活组织带到IMR，以便进行冷冻SEM 现场观察。 我们还希望利用IMR设施， 进行高压冷冻，组织将被进一步处理， 在MTU进行TEM。