LEAP Image Scanning Cytometer/Sorter/Optoinjection Shared Instrument

描述(由申请人提供)：普渡大学发现园宾德利生物科学中心的普渡流式细胞术和细胞分离设备建议购买一台新的LEAP(激光启用分析和处理)(Cynellect，Inc.，San Diego，CA)高速、多色荧光、单细胞、图像扫描/激光光消融/激光光注射设备，用于共享细胞分析/细胞分离设备。这一新上市的仪器不仅提供了高速细胞图像扫描细胞术，还能够通过激光消融去除不需要的贴壁或悬浮细胞(包括潜在的生物危险细胞)进行闭合系统分选，以及一种独特的激光-光注入，它为显微注射和电穿孔提供了一种新的替代方案，用于控制基因或大分子进入异种细胞培养中的特定活细胞。也可以在分泌分子检测的基础上，在单细胞水平上选择/克隆附着的细胞，从而选择高分泌细胞。结合光注入和激光消融分选，可以选择光注入细胞进行进一步生长或进一步加工，例如通过基因表达微阵列分析或蛋白质组分析。PI作为最初的LEAP原型的Beta测试场，并参与了LEAP仪器演变为商业可用仪器的过程，就其功能而言，它远远超过了Beta原型。目前的原型与商业版本如此不同，以至于不再受支持。它也不再能够进行光注入，现在只能在手动激光消融模式下使用。它需要被当前最先进的商业系统取代，该系统具有全自动光注入和消融能力。一个由9个主要用户和3个次要用户组成的小组，由13个国家卫生研究院R01拨款资助，希望通过购买新的LEAP仪器来加强其研究。应用包括上述所有一般领域的使用。用户费用将收回与供应商签订的年度维护合同的成本。将定期举办培训课程，以培训每个主要和次要用户组中的用户，并在未来增加新的主要和次要用户。PI实验室使用测试版原型的经验，以及最近访问Cynellect，Inc.时对商用版的直接经验表明，LEAP仪器通常比流式细胞仪/细胞分选仪操作容易得多。在市面上可用的LEAP仪器中，通过在新的、更直观的图形用户界面背后使用复杂的软件系统，这种易用性得到了极大的增强。但PI的实验室也将提供相当多的专业知识，包括一名训练有素的技术人员，随时准备为每个用户提供帮助或运行样本。

DESCRIPTION (provided by applicant): The Purdue Flow Cytometry and Cell Separation Facility of the Bindley Biosciences Center in Discovery Park at Purdue University proposes to purchase a new LEAP (Laser Enabled Analysis and Processing) (Cyntellect, Inc., San Diego, CA) high-speed, multicolor fluorescence, single cell, image scanning/laser optoablation/ laser optoinjection instrument for a shared cytometry/cell separation facility. This newly commercially available instrumentation not only provides high-speed cell image scanning cytometry, but also the ability to perform closed system sorting of undesired adherent or suspension cells (including potentially biohazardous cells) by laser ablation removal, and a unique laser-optoinjection which provides a new alternative to micro-injection and electroporation for controlled entry of genes or macromolecules into specific living cells in a heterogeneous cell cultures. Attached cells can also be selected/cloned at the single cell level on the basis of secreted molecule assays, allowing selection of high-secreting cells. Combination of optoinjection followed by laser ablation sorting allows selection of optoinjected cells for further growth or for further processing, e.g. by gene expression microarray analysis or proteomic analysis. The PI served as a beta test site for the original LEAP prototype and has been involved in the evolution of the LEAP instrument into a commercially available instrument which goes far beyond the beta prototype in terms of its capabilities. The current prototype is so different from the commercial version that it is no longer supportable. It is also no longer capable of optoinjections and can now only be used in a manual laser ablation mode. It needs to be replaced with a current state-of-the-art commercial system with full automated optoinjection and ablation capabilities. A group of 9 major users and 3 minor users, with 13 funded NIH R01 grants, wishes to enhance its research by acquisition of a new LEAP instrument. Applications include use of all of the above general areas. User fees will recover the costs of yearly maintenance contracts with the vendor. Training sessions will be held periodically to train users within each major and minor group of users, as well as to add new major and minor users in the future. Experience in the PI's laboratory with the beta prototype, and more recently based on direct experience with the commercial version on visits to Cyntellect, Inc. has shown that the LEAP instrument is generally far easier to operate than flow cytometer/ cell sorters. This ease of use has been greatly enhanced in the commercially available LEAP instrument through a sophisticated software system behind a new, much more intuitive, graphical user interface. But there will also be a considerable expertise available from the PI's lab including a trained technician ready to either assist or run samples for each user.