Live-cell, deep-tissue, low-light, 3D-STED confocal microscopy: a super-resolution imaging platform specifically designed for plant science.

活细胞、深层组织、低光、3D-STED 共焦显微镜：专为植物科学设计的超分辨率成像平台。

• 批准号: BB/W019752/1
• 负责人: Alison Roberts
• 金额: $ 103.33万
• 依托单位: James Hutton Institute
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FW019752%2F1
• 关键词: Live; cell; deep; tissue; low

Microscopy is one of the oldest methods used to study complex biological processes and has seen major developments since the creation of the first microscopes in the late 16th century. Fluorescence microscopy has allowed many breakthrough discoveries by giving scientists the ability to locate and track specific molecules within cells. All light microscopes are limited by the physical law of diffraction. However, the development of super resolution (SR)microscopy, which was recognized by a Nobel prize to E.Betzig, S. Hell and E. Moerner in 2014, has revolutionised the range of imaging that can be achieved and biological questions that can be answered. After two decades of combined improvements in these technologies and in associated computing power, SR microscopes are becoming more accessible and can now be customised for individual biological systems. The vast majority of the current super resolution systems have been designed and calibrated by their manufacturers for studies using human and animal cells. However, plant cells are larger and more complex than animal cells, contain thick cell walls and emit significant auto-fluorescence across the visible spectrum. This makes SR imaging extremely challenging in live plant cells and, consequently, few manufacturers have designed microscopes suitable for this purpose. In turn, this lack of SR imaging of plant cells in the UK is impeding breakthrough discoveries and ultimately restricting plant science. The James Hutton Institute has world-renowned expertise in plant and agricultural research. One area of research is to develop crops that are more resilient to climate change and resistant to pests and diseases; subjects that require an understanding of cellular processes to help mitigate risks to plant health and agriculture. Our existing imaging facility has been a core component in our cell and molecular research but advances in microscope technology mean that we need access to super-resolution microscopy techniques to remain at the forefront of cell biology research.We propose to purchase a super-resolution microscope, providing the science community in Scotland and the UK with a powerful system optimised for use in plants. We have found that the STED technology provided by the Abberior Facility-Line 3D-STED microscope was by far the best equipment in recent tests, allowing imaging of both fixed and live plant samples and providing up to 50nm resolution. At present there are no similar microscopes in the UK that are optimised for plant cell imaging, and legislation and plant health regulations restrict us from taking many of our samples off-site to utilise other facilities. Since we have excellent (and licensed) plant growth facilities at JHI it makes sense to install a SR microscope in our existing facility and make it available to the wider biological research community across Tayside and the UK. In time the equipment will be moved to the Advanced Plant Growth Centre; a purpose-built, world-class plant phenotyping facility which is currently being built on the JHI site. Both Dundee and St Andrews Universities are partners in this bid and many researchers (from a wide range of disciplines) have submitted requests to use this resource if it becomes available, covering projects in the areas of sustainable agriculture and food, understanding the rules of life, an integrated understanding of health and transformative technologies. Researchers who receive training to use these techniques and equipment will benefit from continuing professional development, the facility and its core staff will benefit from exposure to new technology and expansion and diversification of our user group, the BBSRC will benefit from enhanced research outputs and end-users will benefit from the outcomes of the research.

显微镜是用于研究复杂生物过程的最古老的方法之一，自世纪末第一台显微镜问世以来，显微镜已经取得了重大发展。荧光显微镜使科学家能够定位和跟踪细胞内的特定分子，从而实现了许多突破性的发现。所有的光学显微镜都受到衍射物理定律的限制。然而，获得诺贝尔奖的E.Betzig，S。地狱和E。Moerner在2014年，彻底改变了可以实现的成像范围和可以回答的生物学问题。经过二十年的综合改进，这些技术和相关的计算能力，SR显微镜变得越来越容易获得，现在可以为单个生物系统定制。目前绝大多数超分辨率系统都是由制造商设计和校准的，用于使用人类和动物细胞的研究。然而，植物细胞比动物细胞更大，更复杂，包含厚的细胞壁，并在可见光谱范围内发出显着的自体荧光。这使得SR成像在活的植物细胞中极具挑战性，因此，很少有制造商设计适合此目的的显微镜。反过来，英国缺乏植物细胞的SR成像阻碍了突破性发现，并最终限制了植物科学。詹姆斯赫顿研究所在植物和农业研究方面拥有世界知名的专业知识。研究的一个领域是开发更能适应气候变化和抵抗病虫害的作物;需要了解细胞过程以帮助减轻植物健康和农业风险的主题。我们现有的成像设备一直是我们细胞和分子研究的核心组成部分，但显微镜技术的进步意味着我们需要获得超分辨率显微镜技术，以保持在细胞生物学研究的前沿。我们建议购买一台超分辨率显微镜，为苏格兰和英国的科学界提供一个强大的系统，优化用于植物。我们发现，Abberior Facility-Line 3D-STED显微镜提供的STED技术是目前为止最好的设备，可以对固定和活体植物样品进行成像，并提供高达50 nm的分辨率。目前，英国还没有针对植物细胞成像进行优化的类似显微镜，立法和植物健康法规限制我们将许多样品带到其他设施。由于我们在JHI拥有优秀的（和许可的）植物生长设施，因此在我们现有的设施中安装SR显微镜并将其提供给泰赛德和英国更广泛的生物研究社区是有意义的。随着时间的推移，设备将被转移到先进的植物生长中心，一个专门建造的，世界级的植物表型分析设施，目前正在建设的JHI网站。邓迪大学和圣安德鲁斯大学都是此次竞标的合作伙伴，许多研究人员（来自广泛的学科）已经提交了使用此资源的请求，如果可用，涵盖可持续农业和食品领域的项目，了解生活规则，对健康和变革技术的综合理解。接受培训使用这些技术和设备的研究人员将受益于持续的专业发展，该设施及其核心工作人员将受益于接触新技术以及我们用户群的扩展和多样化，BBSRC将受益于增强的研究成果，最终用户将受益于研究成果。