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

从混合物中分析特定细胞的能力是许多生物学尖端实验的基础，从罕见的致癌细胞的特征到分离用于化肥的聚磷藻菌株。流式细胞仪(FC)技术彻底改变了我们表征单个细胞的能力。它包括用染料标记细胞，当受到特定波长的光照射时，这些染料会发出光(这个过程称为荧光)，例如通过将染料标记的抗体结合到细胞上。这些标记的细胞随后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标记的细胞的数量，还可以确定标记的强度。通常，细胞被许多不同的染料标记，每种染料都被不同波长的光激活，因此可以根据存在的染料的组合来进一步细分细胞群体。事实上，特定的细胞群可以通过偏转不同标记的细胞来“分类”成不同的种群来分离。自1960年代S发明以来，FC所使用的染料和机器变得越来越敏感和复杂，能够检测到非常罕见的细胞，也允许在无菌条件下将这些罕见的细胞分离为单个细胞。事实上，目前的尖端实验需要能够检测到广泛的染料，以及新开发的用于检测稀有细胞的“明亮”标签。由于机器和内置激光器的年龄，这样的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建立一个集成的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所能进行的实验的复杂性发生重大变化。具体地说，所要求的细胞分析仪可以同时检测13种不同的染料，从而能够检测非常稀有或“昏暗”的细胞，同时细胞分选仪不仅有额外的激光器，这对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对细胞进行分类，这是进行尖端基因编辑技术所必需的。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。这项研究的范围包括对细胞生物学基础研究的分析，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，大脑细胞如何退化，心脏细胞如何在心脏病中衰竭。这些工具还将有助于了解传染病以及如何抗击这些疾病。这些后一项研究调查了一些重要的病毒，如2001年摧毁英国农业的口蹄疫病毒，以及疟疾等人类寄生虫感染。此外，细菌新药的开发和细菌对抗生素的抗药性机制也将受益。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，这些实验范围从筛选藻类吸收磷酸盐用作新肥料的能力，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多名研究人员和他们的相关研究项目将直接受益于这一新设备。

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.