Multi-Laser Flow Cytometer for Polychromatic Analysis

描述（由申请人提供）：我们申请资金购买一台Becton Dickinson定制多激光LSRII流式细胞仪，用于多色流式细胞术分析。范德比尔特大学目前的设施为进行高技术流式细胞术工作提供了强大的基础设施，但目前还不能满足美国国立卫生研究院资助的科学家社区的科学需求。首先，用户组中的许多科学家已经扩大了他们对新荧光团、量子点和变体GFP分子的使用，以加强工作的多色性质，这是复杂功能读出所需要的。许多研究人员在显微镜中使用这些新颖的荧光团组合，但现在需要流式细胞法提供的更定量的分析。目前可用的仪器在进行这些研究的关键光谱区域缺乏激光器。此外，研究人员使用分析仪器的机会越来越有限，因为目前每台仪器的使用时间超过每周45小时，这还不包括每天设置和校准所需的时间。访问的限制已经开始限制我们nih资助的研究人员的进展。该仪器将支持来自医学院9个研究小组的19个主要用户组成的高度协作小组。由主要用户资助的正在进行的项目将受益于疫苗反应、癌症、自身免疫性疾病、发育、基本免疫反应等方面的研究。对于每个项目，所要求的仪器将通过提供所研究细胞的独特的新分析能力，并增加分析细胞术的使用，从而产生直接和实质性的影响。范德比尔特流式细胞术核心实验室（Vanderbilt Flow Cytometry Core Laboratory）是一项机构共享资源，该实验室的收购和操作将保证用户培训/支持、每天24小时、每周7天的访问以及仪器维护和维修。用户费用将包括员工支持、服务和维修。该机构已作出特别强有力的财政承诺，支持该工具的维护和使用。其他研究人员将能够利用现有的时间，因为该仪器将放置在一个机构的核心，从而最大限度地利用和影响。该核心目前支持超过200名用户，我们在范德比尔特接受了分析和分选细胞术方面的培训。

DESCRIPTION (provided by applicant): We request funds to purchase a Becton Dickinson custom multilaser LSRII flow cytometer for polychromatic analytical flow cytometry. The current facilities at Vanderbilt University provide a strong infrastructure for conduct of highly technical flow cytometric work, but do not currently meet the scientific needs for the NIH-funded community of scientists. First, many of the scientists in the user group have expanded their use of new fluorophores, quantum dots, and variant GFP molecules in an effort to intensify the polychromatic nature of the work, which is needed for complex functional readouts. Many of these investigators are using these novel combinations of fluorophores in microscopy, but now need the more quantitative analysis that is afforded by flow cyto- metry. The currently available instruments are lacking lasers in critical regions of the spectrum for the conduct of these studies. In addition, researchers are becoming limited in access to analytical instruments because the amount of use of the current instruments exceeds 45 hours per week per instrument, not counting the time needed for daily setup and calibration. Limitation of access has begun to constrain the progress of our NIH-funded researchers. This instrument will support a highly collaborative group of 19 major users from 9 research groups in the School of Medicine. Ongoing, NIH-funded projects by major users that will benefit include research on vaccine responses, cancer, autoimmunse diseases, development, basic immune responses and others. For each project, the requested instrumentation will have an immediate and substantial impact by providing unique new analytical capabilities of the cells under study, and increased access to analytical cytometry. Acquisition and operation of this cytometer by the Vanderbilt Flow Cytometry Core Laboratory, an institutional shared resource, will guarantee user training/support, 24 hour/day 7 day/week access, and instrument maintenance and repair. User fees will cover staff support, service and repair. The institution has made an exceptionally strong financial commitment to support maintenance and use of the instrument. Other researchers will be able to use available time, as the instrument will be sited in an institutional core, thus maximizing use and impact. This Core currently supports over 200 users who we have trained in analytical and sorting cytometry at Vanderbilt. PUBLIC HEALTH RELEVANCE: The proposed state-of-the-art instrument will give researchers a new capability and expanded access to study important functions of cells including the immune system, stem cells, and cell leading to cancer, thereby improving our ability to develop new targets for intervention in human diseases.