A state-of-the-art flow cytometer for a multi-disciplinary integrative biology department

适用于多学科综合生物学部门的最先进的流式细胞仪

• 批准号: BB/F011180/1
• 负责人: Rachel Lawrence
• 金额: $ 18.62万
• 依托单位: Royal Veterinary College
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2008
• 资助国家: 英国
• 起止时间: 2008 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FF011180%2F1
• 关键词: state; art; flow; cytometer; multi

A Flow Cytometer is a central tool for life science research laboratories. This proposal encompasses projects from three of the four College research themes, all of which currently make use of flow cytometric techniques. They range from the fields of immunology, virology and cardio-vascular and inflammation biology to reproduction and development. We anticipate that should this proposal be funded, the equipment will also be used to collect and analyse data for a number of other projects. A flow cytometer uses a laser and a series of deflecting mirrors to discriminate between different populations of cells and microparticles either by detecting physical differences in the cells and/or by detecting fluorescent markers that can attach to molecules inside or on the surface of the cell. This technique is vital for enumerating specific cell types either in the blood or at the site of disease or, for example, to validate the success of introduction of foreign genes into cells. Due to the wide range of applications of this technique in order for our research to remain at the international forefront of science we require day-to-day use of a flow cytometer that is both easy-to-use and is at the cutting-edge of technology. A flow cytometer has been in use for many years at the RVC and has proved invaluable. However, recent advances in laser technology and the production of new fluorescent dyes and markers mean that our current flow cytometer, a Beckman Coulter EPICS XL-MCL, cannot perform the sophisticated analysis that many current research projects require. For example, our current machine can only detect four colours and analyses 1000 events per second. In addition, a significant expansion in academic staff at the College who undertake immunological research has meant the demand for this technology has dramatically increased. The machine we propose to purchase is the FACS Canto II from Becton Dickinson. In contrast to our EPICS, the FACS Canto II can detect eight separate colours and analyse events at a rate of 10,000 per second. The sensitivity of the proposed flow cytometer is also greatly increased compared with our current machine and enables the measurement and analysis of rare populations of cells in samples taken from human and animals. For example, both eosinophils and T regulatory cells (1-4 % and 3-8% of total white blood cells respectively) which are the subject of current research in the laboratories of Drs Lawrence and Garden respectively, can easily be detected with this instrument. It will also enable a wide range of users to undertake sophisticated studies including detailed and quantifiable analysis of cell growth and cell death and analysis of soluble and cell associated molecules that are produced by cells during infection and chronic diseases in a single small sample volume. Another major advantage of this machine is that it will allow more efficient use of research workers time as not only is the rate of detection ten times faster than the EPICS but acquisition of events by the FACS Canto II can be automated. The more powerful software means that the analysis of data collected will be more efficient and accurate. The RVC places particular emphasis on students gaining laboratory experience and therefore the co-applicants currently host at least 30 students per annum from our BSc, BVetMed (intercalating or elective projects) and MSc courses, as well as supervising approximately 20 PhD students between them at any one time. In addition the RVC, as part of its four year doctoral training programme, has recently instituted a programme of postgraduate training in specific laboratory techniques which is hosted by postdoctoral scientists. The application and practical experience of flow cytometric techniques will be incorporated into this programme. Thus a large number of students will be exposed to this cutting-edge technology whilst undertaking practical laboratory based projects as part of their degree.

流式细胞仪是生命科学研究实验室的核心工具。本提案涵盖了学院四个研究主题中的三个项目，所有这些项目目前都使用流式细胞术技术。它们的范围从免疫学、病毒学、心血管和炎症生物学到生殖和发育。我们预计，如果这项建议得到资助，这些设备也将用于收集和分析其他一些项目的数据。流式细胞仪使用激光和一系列偏转镜，通过检测细胞的物理差异和/或检测可以附着在细胞内部或细胞表面分子上的荧光标记，来区分不同的细胞群和微粒。这项技术对于枚举血液中或疾病部位的特定细胞类型至关重要，例如，验证将外源基因引入细胞是否成功。由于该技术的广泛应用，为了使我们的研究保持在国际科学的前沿，我们需要日常使用既易于使用又处于技术前沿的流式细胞仪。流式细胞仪已在RVC使用多年，并已被证明是无价的。然而，激光技术的最新进展以及新型荧光染料和标记物的生产意味着我们目前的流式细胞仪（Beckman Coulter EPICS XL-MCL）无法执行许多当前研究项目所需的复杂分析。例如，我们目前的机器每秒只能检测四种颜色并分析1000个事件。此外，学院从事免疫学研究的学术人员的显著增加意味着对这项技术的需求急剧增加。我们建议购买的机器是来自Becton Dickinson的FACS Canto II。与我们的EPICS相比，FACS Canto II可以检测到8种不同的颜色，并以每秒10,000个的速度分析事件。与我们目前的机器相比，所提出的流式细胞仪的灵敏度也大大提高，并且能够测量和分析来自人类和动物样本中的稀有细胞群。例如，嗜酸性粒细胞和T调节细胞（分别占白细胞总数的1- 4%和3-8%）是Lawrence博士和Garden博士实验室当前研究的主题，用这种仪器可以很容易地检测到。它还将使广泛的用户能够进行复杂的研究，包括对细胞生长和细胞死亡进行详细和可量化的分析，以及对细胞在感染和慢性疾病期间产生的可溶性分子和细胞相关分子进行分析。这台机器的另一个主要优点是，它将允许更有效地利用研究人员的时间，因为不仅检测速度比EPICS快十倍，而且FACS Canto II的事件采集可以自动化。更强大的软件意味着对收集到的数据的分析将更加高效和准确。RVC特别强调学生获得实验室经验，因此共同申请者目前每年至少接待30名来自我们的理学士，BVetMed（插入或选修项目）和理学硕士课程的学生，以及在任何时候监督大约20名博士生。此外，RVC作为其四年博士培训计划的一部分，最近设立了一个由博士后科学家主持的特定实验室技术研究生培训计划。流式细胞术技术的应用和实践经验将被纳入本课程。因此，大量的学生将接触到这一尖端技术，同时承担实际的实验室项目，作为他们学位的一部分。

项目成果

Filarial parasites develop faster and reproduce earlier in response to host immune effectors that determine filarial life expectancy.

• DOI: 10.1371/journal.pbio.1000525
• 发表时间: 2010-10-19
• 期刊: PLoS biology
• 影响因子: 9.8
• 作者: Babayan SA;Read AF;Lawrence RA;Bain O;Allen JE
• 通讯作者: Allen JE

Eosinophils are important for protection, immunoregulation and pathology during infection with nematode microfilariae.

• DOI: 10.1371/journal.ppat.1003988
• 发表时间: 2014-03
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Cadman ET;Thysse KA;Bearder S;Cheung AY;Johnston AC;Lee JJ;Lawrence RA
• 通讯作者: Lawrence RA

Influence of chylomicron remnants on human monocyte activation in vitro.

• DOI: 10.1016/j.numecd.2010.02.019
• 发表时间: 2011-11
• 期刊: Nutrition, metabolism, and cardiovascular diseases : NMCD
• 作者: Bentley C;Hathaway N;Widdows J;Bejta F;De Pascale C;Avella M;Wheeler-Jones CP;Botham KM;Lawson C
• 通讯作者: Lawson C

The oxidative state of chylomicron remnants influences their modulation of human monocyte activation.

• DOI: 10.1155/2012/942512
• 发表时间: 2012
• 期刊: International journal of vascular medicine
• 影响因子: 1.3
• 作者: Armengol Lopez S;Botham KM;Lawson C
• 通讯作者: Lawson C