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.

生物材料的二维和三维（层析成像）图像提供了对亚细胞结构和功能之间关系的深入了解。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大且非常昂贵。此外，样品制备和/或成像缓慢，阻碍了对生物学研究通常需要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高速度对生物样品进行成像和定量描述，从而分析目前被认为难以进行高通量研究的性状。虽然激光和成像技术已经存在，但要使它们协同工作，关键是要实现生物样品的高分辨率亚细胞成像，还需要进行修改、早期概念探索和验证调查。我们对现有的第一代激光消融（高功率、超快激光脉冲）技术提出了新的改进，使人们能够对以前使用传统技术难以解决的生物问题有新的生物学见解和理解。“biolaser：通过激光消融断层成像进行生物研究”是一个汇集世界级激光技术（剑桥大学制造研究所）、植物研究、生理学（剑桥大学NIAB）和图像分析（两个中心）专业知识的泵浦启动项目。该方法的核心是使用精密激光（基于一种称为“激光消融断层扫描”的技术，简称LAT）。这将与过程监测相结合，根据样品的成分和地形（“计量”）调整烧蚀过程，为显微镜和“化学成像”平台生成前所未有质量的精确烧蚀生物样品，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。使用一系列植物组织，我们将评估和验证设计高精度LAT生物成像系统所需的参数。该项目是围绕关键方法构建的，所有这些方法都必须解决，以成功开发高分辨率生物成像的基于lr的方法：激光制造技术方面的专业知识：国际货币基金组织为该项目提供了世界一流的精密激光研究、工程和制造方面的专业知识。这种现场专业知识是项目成功的基础。精确的烧蚀控制：为了实现高分辨率成像，需要一个均匀的平面。这需要精确了解激光的位置和功率，以及每次激光脉冲后对烧蚀表面的测量，通过过程计量提供，以确保真正实现高分辨率成像。先进的成像和分析工具：生产的精密机械样品将使用一套最先进的2D和3D成像和分析方法进行调查，利用UofC.4的成像专业知识。图像验证：要开发的工具和方法是强大而精确的。然而，在没有参数化和验证的情况下使用，得到的图像可能具有有限的科学价值。生成的图像数据将使用常规方法进行验证，为参数优化提供反馈循环。该项目将为未来通过IFM交付的台式样机制造奠定基础。实现该项目的目标将导致第一代LAT系统的逐步改变，目前工作在低倍率下。最终，我们的目标是在剑桥建立一个LAT生物成像中心，并促进英国和其他地区类似平台的生产。这种能力建设方法将促进充分实现精确LAT生物成像的潜在影响所必需的临界质量技术的采用。

项目成果

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

• 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: Ereful, Nelzo C.;Jones, Huw;Fradgley, Nick;Boyd, Lesley;Cherie, Hirut Assaye;Milner, Matthew J.
• 通讯作者: Milner, Matthew J.

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