Integrating confocal microscopy and cryo plasma FIB milling for tomography

集成共焦显微镜和冷冻等离子体 FIB 铣削用于断层扫描

• 批准号: EP/T033614/1
• 负责人: James Naismith
• 金额: $ 152.9万
• 依托单位: Rosalind Franklin Institute
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FT033614%2F1
• 关键词: Integrating; confocal; microscopy; cryo; plasma

The Life Sciences sector forms a key part of the UK economy: it employs over 220,000 people, contributes significantly toGDP and UK balance of trade, and is crucial for developing leading-edge treatments for patients. It is underpinned by the UK's world-leading research base in the health and life sciences. Many key research breakthroughs are, in turn, enabled by advances in engineering and physical sciences (EPS) research - which provide ever more sophisticated instrumentation and methods to support the study of living organisms (from microbes to plants, animals and the human body) and biological processes (including both disease pathology and drug action). R&D across all parts of this ecosystem - from fundamental understanding to applied research to product development - is crucial for the delivery of long-term economic growth and continued advances in agriculture, food security, healthcare and public health. Historic models of innovation have often been linear, involving a degree of serendipity. Disruptive technologies and scientific breakthroughs will be accelerated if physical scientists, engineers, life scientists and industry work together, and at scale. The Franklin has a focal point (Hub) at Harwell Science and Innovation Campus, linked to formal Spokes in leading HEIs across the UK, it will integrate complementary expertise from academia and industry to create a national centre of excellence for methods development at the convergence of the physical and life sciences.It will create high-value jobs, protect and attract inward investment, and drive long-term growth; and contribute to the delivery of the Government's innovation, industrial and regional strategies.The correlation of visible microscopy with electron imaging, is of course a critical goal of the RFI, it underpins imaging across scales. Thus for cells and tissue, the accurate targeting of the zone of interest to be milled is critical. Due to the crowded cellular environment, it is, as a practical matter, almost impossible to confirm the region of interest is within the EM volume during measurement, since the sample is damaged by electron dose. Correlative Light and EM (CLEM) methods are used to guide the milling and imaging. In broad terms fluorescent tag(s) are put onto the protein(s) of interest; this is extremely technically challenging and prone to error. We propose, in a collaboration with Thermo Fischer Scientific , to combine fluorescence imaging and plasma FIB milling in a single machine, thus transforming the workflow for tomography. This equipment has applications in both correlative imaging themes and in the mass spectrometry theme.

生命科学部门是英国经济的重要组成部分：它拥有超过220，000名员工，为GDP和英国贸易平衡做出了重大贡献，并且对于为患者开发尖端治疗至关重要。它以英国在健康和生命科学领域的世界领先研究基地为基础。许多关键的研究突破反过来又得益于工程和物理科学（EPS）研究的进步，这些研究提供了越来越复杂的仪器和方法，以支持对活生物体（从微生物到植物，动物和人体）和生物过程（包括疾病病理学和药物作用）的研究。这一生态系统的所有部分的研发-从基础知识到应用研究再到产品开发-对于实现长期经济增长以及农业、粮食安全、医疗保健和公共卫生的持续进步至关重要。历史上的创新模式往往是线性的，涉及一定程度的偶然性。如果物理科学家、工程师、生命科学家和工业界共同努力，并形成规模，颠覆性技术和科学突破将加速发展。富兰克林在哈威尔科学与创新园区设有一个联络点（枢纽），与英国各地领先的高等院校的正式辐条相连，它将整合学术界和工业界的互补专业知识，创建一个物理科学和生命科学融合的国家卓越方法开发中心。它将创造高价值的就业机会，保护和吸引外来投资，并推动长期增长;可见光显微镜与电子成像的相关性当然是RFI的一个关键目标，它支持跨尺度成像。因此，对于细胞和组织而言，精确靶向待研磨的感兴趣区域至关重要。由于拥挤的细胞环境，实际上几乎不可能在测量期间确认感兴趣区域在EM体积内，因为样品被电子剂量损坏。相关光和EM（CLEM）方法用于指导铣削和成像。从广义上讲，将荧光标签置于感兴趣的蛋白质上;这在技术上极具挑战性，并且容易出错。我们建议与Thermo Fischer Scientific合作，将联合收割机荧光成像和等离子FIB研磨结合在一台机器中，从而改变断层扫描的工作流程。该设备在相关成像主题和质谱主题中都有应用。