A Multi-User Flow Cytometry Facility for the Biosciences in Leeds

利兹生物科学多用户流式细胞仪设施

• 批准号: BB/R000352/1
• 负责人: George Blair
• 金额: $ 23.98万
• 依托单位: University of Leeds
• 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%2FR000352%2F1
• 关键词: Multi; User; Flow; Cytometry; Facility

The ability to analyse specific cells from a mixture is fundamental to numerous cutting edge experiments in biology, ranging from the characterization of rare cancer causing cells to the isolation phosphate-accumulating strains of algae for use in fertilisers. The technique of flow cytometry (FC) has revolutionized our ability to characterise individual cells. It involves labelling cells with dyes that emit light (the process called fluorescence) when irradiated with light of defined wavelength, for example by binding dye-labelled antibodies to cells. These labelled cells are then detected by sophisticated fluorescence-activated cell analysers, using lasers to determine not only the number of dye-labelled cells but also the intensity of the labelling. Often cells are labelled with a number of different dyes, each activated by different wavelengths of light, and thus it is possible to further sub-divide populations of cells based on the combinations of dyes present. Indeed specific populations of cells can be isolated by deflecting the differently labelled cells to "sort" them into distinct populations.Since its development in the 1960's, the dyes and machines used for FC have become increasingly sensitive and sophisticated, allowing very rare cells to be detected and also allowing these rare cells to be isolated as single cells, under sterile conditions. Indeed, current cutting-edge experiments require the ability to detect a broad spectrum of dyes as well as the newly developed "bright" labels to detect rare cells. Such experiments are not currently possible in Leeds due to the age of the machines and their inbuilt lasers. Here, we request state-of-the-art machines to build an integrated FC facility to address a series of fundamental questions in cell biology, infectious diseases and biotechnology. It is anticipated that these machines will precipitate a step-change in the sophistication of the experiments that we can perform. Specifically, the cell analyser requested can detect 13 different dyes simultaneously, allowing detection of very rare or "dim" cells whilst the cell sorter not only has an additional laser that is critical for detection of a very common red dye, but can also reliably sort cells under sterile conditions, which is essential to carry out cutting edge gene editing techniques.These instruments will give momentum to a plethora of experiments, encompassing research groups from three different Faculties of Leeds University. This research ranges from the analysis of basic studies on the biology of cells, including how cancer cells form and develop, how the precursors to blood cells (stem cells) develop, how cells in the brain degenerate and how cells in the heart fail in heart disease. The instruments will also aid understanding of infectious diseases and how they can be combatted. These latter studies investigate important viruses like foot-and-mouth-disease virus that devastated UK agriculture in 2001 and human parasite infections such as malaria. Also the development of new medicines from bacteria and the mechanisms of bacterial resistance to antibiotics will benefit. Finally, the instruments requested will be important to a series of economically valuable biotechnology experiments that range from screening algae for their ability to take up phosphate for use as new fertilisers to the development of nanomedicines for cancer treatments. In all, 31 Leeds academics have research projects that rely on this equipment. When their research fellows, students and collaborators are included, over 100 researchers and their associated research projects will directly benefit from this new equipment.

从混合物中分析特定细胞的能力是生物学中众多前沿实验的基础，从罕见致癌细胞的表征到分离用于肥料的藻类聚磷菌株。流式细胞术（FC）技术已经彻底改变了我们对单个细胞的鉴定能力。它涉及用染料标记细胞，当用确定波长的光照射时，这些染料会发光（称为荧光的过程），例如通过将染料标记的抗体结合到细胞上。然后，这些标记的细胞被复杂的荧光激活细胞分析仪检测到，使用激光不仅可以确定染料标记细胞的数量，还可以确定标记的强度。细胞通常被标记有许多不同的染料，每种染料被不同波长的光激活，因此可以基于存在的染料的组合进一步细分细胞群体。事实上，通过将不同标记的细胞偏转，将它们“分选”成不同的细胞群，可以分离出特定的细胞群。自20世纪60年代发展以来，用于FC的染料和机器变得越来越敏感和复杂，可以检测到非常罕见的细胞，也可以在无菌条件下将这些罕见的细胞分离为单细胞。事实上，目前的尖端实验需要检测广谱染料的能力，以及新开发的“明亮”标签来检测稀有细胞。由于机器及其内置激光器的年龄，这种实验目前在利兹是不可能的。在这里，我们要求最先进的机器来建立一个综合的FC设施，以解决细胞生物学，传染病和生物技术中的一系列基本问题。预计这些机器将促使我们可以进行的实验的复杂性发生重大变化。具体而言，所要求的细胞分析仪可以同时检测13种不同的染料，从而可以检测非常罕见或“暗淡”的细胞，而细胞分选仪不仅具有额外的激光器，这对于检测非常常见的红色染料至关重要，而且还可以在无菌条件下可靠地分选细胞，这对于开展尖端的基因编辑技术至关重要。这些仪器将为大量实验提供动力，包括来自利兹大学三个不同学院的研究小组。这项研究的范围从分析细胞生物学的基础研究，包括癌细胞如何形成和发展，血细胞（干细胞）的前体如何发展，大脑中的细胞如何退化以及心脏病中的细胞如何衰竭。这些仪器还将有助于了解传染病以及如何防治传染病。这些研究调查了重要的病毒，如2001年摧毁英国农业的口蹄疫病毒和人类寄生虫感染，如疟疾。从细菌中开发新药和细菌对抗生素的耐药性机制也将受益。最后，所要求的仪器将对一系列具有经济价值的生物技术实验至关重要，这些实验的范围从筛选藻类吸收磷酸盐作为新肥料的能力到开发用于癌症治疗的纳米药物。总共有31名利兹学者的研究项目依赖于这种设备。如果将他们的研究员、学生和合作者包括在内，100多名研究人员及其相关研究项目将直接受益于这一新设备。

项目成果

Persistent Replication of a Chikungunya Virus Replicon in Human Cells Is Associated with Presence of Stable Cytoplasmic Granules Containing Nonstructural Protein 3.

人类细胞中Chikungunya病毒复制子的持续复制与存在含有非结构蛋白3的稳定细胞质颗粒有关。

• DOI: 10.1128/jvi.00477-18
• 发表时间: 2018-08-15
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Remenyi R;Gao Y;Hughes RE;Curd A;Zothner C;Peckham M;Merits A;Harris M
• 通讯作者: Harris M

Internalization of rabies virus glycoprotein differs between pathogenic and attenuated virus strains.

• DOI: 10.1099/jgv.0.001935
• 发表时间: 2023-12
• 期刊: JOURNAL OF GENERAL VIROLOGY
• 影响因子: 3.8
• 作者: Almasoud, Ibrahim;Charlton, Frank W.;Finke, Stefan;Barr, John N.;Mankouri, Jamel
• 通讯作者: Mankouri, Jamel

Cellular sheddases are induced by Merkel cell polyomavirus small tumour antigen to mediate cell dissociation and invasiveness.

• DOI: 10.1371/journal.ppat.1007276
• 发表时间: 2018-09
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Nwogu N;Boyne JR;Dobson SJ;Poterlowicz K;Blair GE;Macdonald A;Mankouri J;Whitehouse A
• 通讯作者: Whitehouse A

Dysregulation of the miR-30c/DLL4 axis by circHIPK3 is essential for KSHV lytic replication.

• DOI: 10.15252/embr.202154117
• 发表时间: 2022-05-04
• 期刊: EMBO REPORTS
• 影响因子: 7.7
• 作者: Harper, Katherine L.;Mottram, Timothy J.;Anene, Chinedu A.;Foster, Becky;Patterson, Molly R.;McDonnell, Euan;Macdonald, Andrew;Westhead, David;Whitehouse, Adrian
• 通讯作者: Whitehouse, Adrian

Effective delivery and selective insecticidal activity of double-stranded RNA via complexation with diblock copolymer varies with polymer block composition.

双链RNA通过与二嵌段共聚物络合的有效递送和选择性杀虫活性随聚合物嵌段组成而变化。

• DOI: 10.1002/ps.7793
• 发表时间: 2023
• 期刊: Pest management science
• 影响因子: 4.1
• 作者: Pugsley CE