MoFlo to MoFlo XDP Upgrade for BSL3 Sorting

描述(由申请人提供)：这项申请满足了约翰霍普金斯大学(JHU)对感染了对人类有生物危害的有机体的样本进行高速细胞分选的持续需求。它请求支持将位于约翰霍普金斯大学彭博公共卫生学院生物安全3级(BSL3)实验室的Beckman Coulter MoFlo Legacy流式细胞仪升级为功能相当于MoFlo XDP流式细胞仪的仪器。我们讨论的MoFlo Legacy细胞仪已经有11年多的历史了，作为支持大量依赖它的研究项目(特别是在艾滋病毒/艾滋病、癌症、神经退行性疾病和其他适合在受保护环境中分选的人类细胞疾病领域的项目)的关键工具，它很脆弱，自2013年底起将不再得到制造商的支持。由于JHU的大多数细胞分选设施指的是我们对可能含有肝炎病毒或其他病原体等病原体的人类样本进行分选，而我们的实验室是JHU的主要生物安全3级(BSL3)细胞分选设施，因此可靠和足够复杂的分选器对于该设施的持续成功至关重要，自获得当前的细胞仪以来，该分选器已为至少70份出版物提供了数据，包括在了解和护理艾滋病毒感染方面的重要进展。该实验室团队在管理第三核电站核心设施方面经验丰富，本申请的首席调查员自以来一直指导该设施。随着目前的MoFlo细胞仪已经达到其预期寿命，升级和现代化是a)维持和扩大目前的研究规模所必需的，以及b)为此比购买新的分选细胞仪更具成本效益。在依赖这一设施的艾滋病毒相关研究领域中，包括了解体内艾滋病毒储存库的产生和可能的机制，艾滋病毒感染的免疫发病机制，以及艾滋病毒相关感染的发病机制。人类研究的其他领域包括神经系统疾病的发病机制，干细胞在癌症和其他疾病中的作用，以及许多人类疾病的基因调控方面。介绍了5个主要调查人员和许多其他核心用户的具体研究项目，这些项目将受益于该设施的持续可获得性和可靠性，以及MoFlo升级的具体特点，以证明其收购的合理性。许多其他项目也将在其整个生命周期内使用新的分拣机。描述了维持细胞分选设施未来支持JHU艾滋病毒和人类研究能力的程序和计划，以及该设施运营的财务计划，包括机构支持，包括支付升级后的细胞仪的服务合同和可能出现的任何资金短缺。该设施将由一名经验丰富的全职细胞仪操作员和一名新操作员操作，由设施主任进行培训和监督，并由内部咨询委员会监督。随着要求的升级，该设施将继续在多年内为艾滋病毒感染和其他人类疾病的治疗、管理和预防做出独特贡献。

DESCRIPTION (provided by applicant): This application addresses the ongoing need at Johns Hopkins University (JHU) for high-speed cell sorting of specimens infected with organisms which are biohazardous to humans. It requests support to upgrade a Beckman Coulter MoFlo Legacy flow cytometer, housed in a biosafety level 3 (BSL3) laboratory in the Johns Hopkins Bloomberg School of Public Health, to the functional equivalent of a MoFlo XDP flow cytometer. The MoFlo Legacy cytometer in question is more than 11 years old, is tenuous as a critical instrument to support a large number of research projects that depend on it (specifically projects in the areas of HIV/AIDS, cancer, neurodegenerative diseases, and other diseases of human cells where sorting in a protected environment is appropriate), and will no longer be supported by the manufacturer as of late 2013. As most cell sorting facilities at JHU refer to us the sorting of human samples that may contain agents such as hepatitis viruses or other pathogens, and our laboratory is the main biosafety level 3 (BSL3) cell sorting facility at JHU, a reliable and adequately sophisticated sorter is critical for the continued success of this facility, which has contributed data to at least 70 publications, including important advances in understanding and care of HIV infection, since the acquisition of the current cytometer. The laboratory team is well experienced in management of a BSL3 core facility, with the principal investigator of this application having directed this facility since 1989. With the current MoFlo cytometer now reaching its life expectancy, upgrading and modernizing it are a) needed to sustain and expand the current scale of research, and b) much more cost-effective for this purpose than purchasing a new sorting cytometer. Among the HIV-related research areas that depend on this facility are understanding the generation and possible mechanisms of eradication of HIV reservoirs in vivo, immune pathogenesis of HIV infection, and pathogenesis of HIV-related infections. Other areas of human investigation include pathogenesis of neurological disease, role of stem cells in cancer and other diseases, and aspects of gene regulation in many human diseases. Specific research projects of 5 major investigators and numerous other users of the core that would benefit from continued availability and reliability of this facility are described, as are the specific features of the MoFlo upgrade that justify its acquisition. Many other projects would also use the new sorter over its lifetime. Procedures and plans for sustaining the cell sorting facility's ability to support HIV and human research at JHU for the future are described, as are financial plans for operation of the facility including institutional support comprised of covering the service contract for the upgraded cytometer and any financial shortfalls that may develop. The facility will be operated by an experienced full-time cytometer operator and a new operator to be trained and supervised by the facility director, with oversight by an internal advisory committee. With the requested upgrade, the facility will continue to make unique contributions to the treatment, management, and prevention of HIV infection and other human diseases for many years.