Quantitative and Real-Time Image Analysis for Advanced Light Microscopy.

先进光学显微镜的定量和实时图像分析。

• 批准号: MR/S005382/1
• 负责人: Dominic Waithe
• 金额: $ 44.06万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Fellowship
• 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=MR%2FS005382%2F1
• 关键词: Quantitative; Real; Time; Image; Analysis

In microscopy based research it is common to acquire and analyze microscopy images of cells sampled from populations which have undergone differential treatments. A non-exhaustive number of samples (i.e. images of cells) are often acquired due to experimental limitations that include: restricted acquisition periods, provision of expensive equipment or pressures to move onto the next experiment. Random sampling is a methodology that is used to sample representatively. There are some good reasons however for not sampling randomly. For example, cells maybe expressing a reporter protein and the copy number (quantity) of this molecule maybe too high or too low for subsequent analysis and so preference is shown toward those examples where the expression levels are within a certain range. Or, alternatively, the cells in an area maybe compromised by an artifact, be unhealthy, or too dense or sparse for a particular analysis. There are many factors that will influence the choices of an imaging researcher that are fully justifiable, but it is a problem as to how this information is documented and shared with other scientists. Removing the scientist from the pipeline of acquisition and analysis is counter-productive, a better solution is to provide tools that allow them to document and describe any experimental subjectivity as well as boosting reproducibility through automation. The objectives of this first part of the project (1-4) revolve around solving these issues of documenting, describing and automating image acquisition so that experimental design can be better communicated between users and laboratories. Furthermore, we would like to develop tools and approaches, which allow better visualization and real-time feedback for advanced approaches so as to better inform imaging scientists as they perform their research and make experimental decisions. The second part of the project relates to this goal, objectives 5-6.

在基于显微镜的研究中，通常获取和分析从经历了差异处理的群体中取样的细胞的显微镜图像。由于实验限制，通常会采集非穷尽数量的样本（即细胞图像），这些实验限制包括：有限的采集周期、提供昂贵的设备或进入下一个实验的压力。随机抽样是一种用于代表性抽样的方法。然而，有一些很好的理由不随机抽样。例如，细胞可能表达报告蛋白，并且该分子的拷贝数（量）对于随后的分析可能太高或太低，因此优选表达水平在一定范围内的那些实施例。或者，可替代地，区域中的细胞可能受到伪影的损害，不健康，或者对于特定分析来说太密集或稀疏。有许多因素会影响成像研究人员的选择，这些因素是完全合理的，但如何记录这些信息并与其他科学家共享是一个问题。将科学家从采集和分析的管道中移除是适得其反的，更好的解决方案是提供工具，允许他们记录和描述任何实验主观性，并通过自动化提高可重复性。该项目第一部分（1-4）的目标围绕着解决这些记录，描述和自动化图像采集的问题，以便用户和实验室之间可以更好地沟通实验设计。此外，我们希望开发工具和方法，为先进的方法提供更好的可视化和实时反馈，以便在成像科学家进行研究和做出实验决策时更好地为他们提供信息。该项目的第二部分涉及这一目标，即目标5-6。

项目成果

Oncogenic Gata1 causes stage-specific megakaryocyte differentiation delay

致癌Gata1导致阶段特异性巨核细胞分化延迟

• DOI: 10.1101/791079
• 发表时间: 2019
• 作者: Juban G

A tissue-specific self-interacting chromatin domain forms independently of enhancer-promoter interactions.

• DOI: 10.1038/s41467-018-06248-4
• 发表时间: 2018-09-21
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Brown JM;Roberts NA;Graham B;Waithe D;Lagerholm C;Telenius JM;De Ornellas S;Oudelaar AM;Scott C;Szczerbal I;Babbs C;Kassouf MT;Hughes JR;Higgs DR;Buckle VJ
• 通讯作者: Buckle VJ

Diffusion and interaction dynamics of the cytosolic peroxisomal import receptor PEX5

胞质过氧化物酶体输入受体 PEX5 的扩散和相互作用动力学

• DOI: 10.1101/2021.05.25.445571
• 发表时间: 2021
• 作者: Galiani S

Challenges of Using Expansion Microscopy for Super-resolved Imaging of Cellular Organelles.

• DOI: 10.1002/cbic.202000571
• 发表时间: 2021-02-15
• 期刊: Chembiochem : a European journal of chemical biology
• 作者: Büttner M;Lagerholm CB;Waithe D;Galiani S;Schliebs W;Erdmann R;Eggeling C;Reglinski K
• 通讯作者: Reglinski K