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

点击翻译按钮获取中文摘要

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.