Life in 3D, expanding the structural length scales via Serial Block Face Scanning Electron Microscopy (SBF-SEM)

3D 生活，通过串行块面扫描电子显微镜 (SBF-SEM) 扩展结构长度尺度

• 批准号: BB/R01387X/1
• 负责人: Paul Verkade
• 金额: $ 76.45万
• 依托单位: University of Bristol
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FR01387X%2F1
• 关键词: Life; 3D; expanding; structural; length

Bioimaging is one of the key technologies for life science research but the way life functions has however often been studied by reducing this to 2-dimensional projections for instance by light and / or electron microscopy. Although this approach has led and will also in the future lead to breakthrough discoveries there is a realisation that if possible "life" should be studied as much as possible in a 3D environment, ideally in the context of a tissue or organism. Some of the newer microscopy techniques therefore are focussing on this aspect and allow for imaging large volumes in all 3 dimensions, e.g. by using light sheet light microscopy. Electron Microscopy has generally focussed on higher resolution structural information but alongside the "resolution revolution" in structural cryo Electron Microscopy (EM) which is revealing ever greater molecular details, there has been a quieter but probably equally important revolution in volume EM. By sequentially scanning the block face of an embedded sample and removing the scanned top layer, a large high-resolution 3-dimensional volume can be acquired. Although at somewhat lower resolution this volume is much larger than possible by standard (serial thick section) Electron Tomography. This allows for the structural visualisation of complete cells and the interactions of these cells with other cells or the extracellular matrix inside a tissue. As all life is built in 3 Dimensions, Serial Block Face Scanning Electron Microscopy (SBF-SEM) has now become the method of choice for a growing number of life science studies.We would like to apply for a Scanning Electron Microscope (SEM) with an integrated diamond knife based microtome. We will integrate the technology into the Wolfson Bioimaging Facility, the centralised microscopy facility of the Faculty of Biomedical Sciences at the University of Bristol which houses both light and electron microscopy and is well-known for its Correlative Microscopy technologies.We will apply the tool to study a variety of research questions that can only be studied in 3D. These include but are certainly not limited to the formation of synapses in the brain, the infiltration of macrophages into tissue, and the formation and release of platelets in the blood stream. Importantly this tool will allow us to also study the interaction of the extracellular matrix with cells, an area where 3D imaging is critical.Last but not least we will use the microscope to advance our understanding in the Synthetic Biology field, one of the highlight areas of the BBSRC. Some of the Synthetic Biology research in Bristol focusses on the development of artificial extracellular supports with the idea to grow cells on, and ultimately to use these for transplantation. In order to study the interaction of seeded cells with such matrix supports the SBFSEM will be of critical importance.Training of the next generation of scientists is an important aspect within the Wolfson Bioimaging Facility. As home of an international EMBO course and other national training courses the technology would be of special interest to seamlessly integrate into some of those training courses, providing students with training in the latest state of the art technology.

生物成像是生命科学研究的关键技术之一，但是生命功能的方式通常通过将其减少到二维投影来研究，例如通过光学和/或电子显微镜。虽然这种方法已经导致并将在未来导致突破性的发现，但人们认识到，如果可能的话，“生命”应该尽可能在3D环境中进行研究，理想情况下是在组织或生物体的背景下。因此，一些较新的显微镜技术集中在这方面，并允许在所有3个维度上成像大体积，例如通过使用光片光学显微镜。电子显微镜通常专注于更高分辨率的结构信息，但随着结构低温电子显微镜（EM）的“分辨率革命”，它揭示了更大的分子细节，在体积EM中有一个安静但可能同样重要的革命。通过顺序扫描包埋样品的块面并去除扫描的顶层，可以获得大的高分辨率三维体积。尽管分辨率稍低，但该体积比标准（连续厚切片）电子断层扫描可能的体积大得多。这允许完整细胞的结构可视化以及这些细胞与组织内的其他细胞或细胞外基质的相互作用。由于所有的生命都是三维的，连续块面扫描电子显微镜（SBF-SEM）现在已经成为越来越多的生命科学研究的首选方法。我们想申请一台带有集成金刚石刀切片机的扫描电子显微镜（SEM）。我们将把这项技术整合到沃尔夫森生物成像设备中，沃尔夫森生物成像设备是布里斯托大学生物医学科学学院的中央显微镜设备，它拥有光学显微镜和电子显微镜，并以其相关显微镜技术而闻名。我们将应用该工具研究各种只能在3D中研究的研究问题。这些包括但当然不限于脑中突触的形成、巨噬细胞向组织中的浸润以及血小板在血流中的形成和释放。重要的是，这一工具还将使我们能够研究细胞外基质与细胞的相互作用，这是3D成像至关重要的领域。最后但并非最不重要的是，我们将使用显微镜来推进我们对合成生物学领域的理解，这是BBSRC的亮点领域之一。布里斯托的一些合成生物学研究集中在人工细胞外支持物的开发上，其想法是在其上生长细胞，并最终将其用于移植。为了研究接种细胞与这种基质支持物的相互作用，SBFSEM将是至关重要的。下一代科学家的培训是沃尔夫森生物成像设施的一个重要方面。作为国际EMBO课程和其他国家培训课程的所在地，该技术将特别感兴趣地无缝集成到其中一些培训课程中，为学生提供最新技术的培训。

项目成果

A cellular and molecular atlas reveals the basis of chytrid development.

细胞和分子图谱揭示了壶菌发育的基础。

• DOI: 10.7554/elife.73933
• 发表时间: 2022-03-01
• 期刊: eLife
• 影响因子: 7.7
• 作者: Laundon D;Chrismas N;Bird K;Thomas S;Mock T;Cunliffe M
• 通讯作者: Cunliffe M

The marine gastropod Conomurex luhuanus (Strombidae) has high-resolution spatial vision and eyes with complex retinas

海洋腹足动物 Conomurex luhuanus（Strombidae）具有高分辨率的空间视觉和具有复杂视网膜的眼睛

• DOI: 10.1101/2021.12.21.473630
• 发表时间: 2021
• 作者: Irwin A