BioLaser: Establishing a High-Resolution Laser Ablation Tomography Platform for UK Bioimaging Research

BioLaser：为英国生物成像研究建立高分辨率激光烧蚀断层扫描平台

• 批准号: BB/P027458/1
• 负责人: Eric Ober
• 金额: $ 17.59万
• 依托单位: National Institute of Agricultural Botany
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FP027458%2F1
• 关键词: BioLaser; Establishing; Resolution; Laser; Ablation

Background summaryTwo- and three-dimensional (tomographic) images of biological materials provide insight into the relationships between subcellular structure and function. However, particularly for hard or opaque tissues, instruments that can do this are currently large and extremely expensive. Furthermore, sample preparation and/or imaging is slow, preventing cost effective analysis of the many samples commonly required for biological studies. Thus, there is urgent need for new methods to image and quantitatively describe biological samples at high resolution and speed, allowing analysis of traits currently considered intractable to high-throughput investigation. While the component laser and imaging technologies exist, getting them to work together, and crucially, to achieve high-resolution sub-cellular imaging on biological samples, requires modifications and early concept exploratory and validation investigations. We propose novel modifications to existing, first-generation laser ablation (high-powered, ultrafast laser pulses) technology, enabling new biological insight and understanding into biological problems that were previously intractable using conventional techniques.Project summary'BioLaser: Biological Investigations by Laser Ablation Tomography' is a pump-priming project that aligns world-class expertise in laser technology (Institute for Manufacturing, IFM, at the University of Cambridge), plant research, physiology (NIAB, Cambridge) and image analysis (both centres). The approach centers around the use of precision lasering (based on a technique called 'Laser Ablation Tomography', LAT). This will be combined with in-process monitoring to adjust the ablation process according to the composition and topography of the sample ('metrology'), to generate precision ablated biological samples of unprecedented quality for microscopy and 'chemical imaging' platforms that render high-resolution 2D & 3D images, allowing quantitative analysis of cellular and subcellular structures. Using an array of plant tissues, we will evaluate & validate the parameters required to design a precision LAT bioimaging system. This project is structured around for key approaches, all of which must be addressed to successfully develop LAT-based methodologies for high-resolution bioimaging:1. Expertise in laser-based manufacturing technologies: IMF provides this project with world-class expertise in precision laser research, engineering and manufacturing. Such on-site expertise is the foundation for project success.2. Precise ablation control: To achieve high-resolution imaging, a uniform plane is required. This needs precise knowledge of the position and power of the laser alongside measurement of the ablated surface after each laser pulse, provided via in-process metrology to ensure high-resolution imaging is truly achievable.3. Advanced tools for imaging and analysis: The precision machined samples produced will be investigated using a suite of state-of-the-art 2D & 3D imaging and analysis approaches, exploiting imaging expertise at the UofC.4. Image validation: the tools and approaches to be developed are powerful and precise. However, used without parameterisation and validation, the resulting images could be of limited scientific value. The image data generated will be validated using conventional approaches, providing a feedback loop for parameter optimisation.This project will lay the foundation for future manufacture of a benchtop prototype, delivered via IFM. Achieving the project aims would result in a step-change over first generation LAT systems that currently work at low magnification. Ultimately, we aim to establish a LAT bioimaging hub based in Cambridge, and to facilitate the production of similar platforms for the UK and beyond. This capacity-building approach will promote the critical-mass in technology uptake necessary for the potential impact of precision LAT bioimaging to be fully realised.

背景摘要生物材料的二维和三维（断层扫描）图像可以深入了解亚细胞结构和功能之间的关系。然而，特别是对于坚硬或不透明的组织，目前能够做到这一点的仪器体积庞大且极其昂贵。此外，样品制备和/或成像速度缓慢，阻碍了对生物学研究通常所需的许多样品进行成本有效的分析。因此，迫切需要新的方法以高分辨率和速度对生物样本进行成像和定量描述，从而能够分析目前被认为难以进行高通量研究的性状。虽然组件激光和成像技术已经存在，但要让它们协同工作，并且最重要的是，要在生物样品上实现高分辨率亚细胞成像，需要进行修改以及早期概念探索和验证研究。我们提出对现有的第一代激光烧蚀（高功率、超快激光脉冲）技术进行新颖的修改，从而实现对以前使用传统技术难以解决的生物问题的新的生物学见解和理解。项目摘要“BioLaser：激光烧蚀的生物学研究” “断层扫描”是一个泵启动项目，结合了激光技术（剑桥大学 IFM 制造研究所）、植物研究、生理学（剑桥 NIAB）和图像分析方面的世界一流专业知识（两个中心）。该方法的核心是使用精密激光（基于一种称为“激光烧蚀断层扫描”（LAT）的技术）。这将与过程中的监测相结合，根据样品的成分和形貌（“计量学”）调整消融过程，为显微镜和“化学成像”平台生成前所未有的质量的精密消融生物样品，从而提供高分辨率 2D 和 3D 图像，允许对细胞和亚细胞结构进行定量分析。我们将使用一系列植物组织来评估和验证设计精密 LAT 生物成像系统所需的参数。该项目围绕关键方法构建，必须解决所有这些问题才能成功开发基于 LAT 的高分辨率生物成像方法：1。激光制造技术方面的专业知识：IMF 为该项目提供了精密激光研究、工程和制造方面的世界一流的专业知识。这种现场专业知识是项目成功的基础。2．精确的烧蚀控制：为了实现高分辨率成像，需要均匀的平面。这需要精确了解激光的位置和功率，同时在每个激光脉冲后测量烧蚀表面，通过过程中计量提供，以确保真正实现高分辨率成像。3。先进的成像和分析工具：将利用 UofC.4 的成像专业知识，使用一套最先进的 2D 和 3D 成像和分析方法对生产的精密加工样品进行研究。图像验证：要开发的工具和方法强大且精确。然而，如果在没有参数化和验证的情况下使用，所得图像的科学价值可能有限。生成的图像数据将使用传统方法进行验证，为参数优化提供反馈回路。该项目将为未来通过 IFM 交付的台式原型制造奠定基础。实现该项目目标将导致目前在低放大倍数下工作的第一代 LAT 系统发生重大变化。最终，我们的目标是在剑桥建立一个 LAT 生物成像中心，并促进为英国及其他地区生产类似的平台。这种能力建设方法将促进充分实现精密 LAT 生物成像的潜在影响所必需的技术采用的临界质量。

项目成果

BioLaser : establishing a high-resolution laser ablation tomography platform for UK bioimaging research

BioLaser：为英国生物成像研究建立高分辨率激光烧蚀断层扫描平台

• 发表时间: 2022
• 作者: Atkin Peter

Nutritional and genetic variation in a core set of Ethiopian Tef (Eragrostis tef) varieties

埃塞俄比亚 Tef（Eragrostis tef）核心品种的营养和遗传变异

• DOI: 10.1186/s12870-022-03595-9
• 发表时间: 2022-04-28
• 期刊: BMC Plant Biology
• 影响因子: 5.3
• 作者: N. Ereful;H. Jones;N. Fradgley;L. Boyd;Hirut Assaye Cherie;Matthew J. Milner
• 通讯作者: Matthew J. Milner