LEAP Image Scanning Cytometer/Sorter/Optoinjection Shared Instrument

描述（由申请人提供）：普渡大学Discovery Park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（激光分析和处理）（Cyntellect, Inc., San Diego， CA）高速，多色荧光，单细胞，图像扫描/激光光消融/激光光注射仪器，用于共享细胞术/细胞分离设备。这种新上市的商业仪器不仅提供高速细胞图像扫描细胞术，而且还能够通过激光消融去除对不需要的贴壁或悬浮细胞（包括潜在的生物危险细胞）进行封闭系统分选，并且独特的激光光注射提供了微注射和电穿孔的新选择，用于控制基因或大分子进入异质细胞培养物中的特定活细胞。在分泌分子测定的基础上，也可以在单细胞水平上选择/克隆附着细胞，从而选择高分泌细胞。光注射与激光消融分选相结合，可以选择光注射细胞进行进一步生长或进一步处理，例如通过基因表达微阵列分析或蛋白质组学分析。PI作为原始LEAP原型的beta测试站点，参与了LEAP仪器向商用仪器的发展，其功能远远超过beta原型。目前的原型机与商业版本差别太大，已不再支持。它也不再能够进行光注射，现在只能在手动激光消融模式下使用。它需要被目前最先进的商业系统所取代，该系统具有全自动光注入和消融功能。一个由9个主要用户和3个次要用户组成的小组，拥有13个NIH R01资助，希望通过获得新的LEAP仪器来加强其研究。应用包括使用所有上述一般领域。用户费用将收回与供应商签订的年度维护合同的成本。将定期举行培训课程，培训每一主要和次要用户组内的用户，并在将来增加新的主要和次要用户。在PI实验室使用beta原型的经验，以及最近访问Cyntellect公司的商业版本的直接经验表明，LEAP仪器通常比流式细胞仪/细胞分选仪更容易操作。这种易用性在商用LEAP仪器中得到了极大的增强，通过一个复杂的软件系统，背后是一个新的，更直观的图形用户界面。但PI的实验室也将有相当多的专业知识，包括训练有素的技术人员准备协助或为每个用户运行样本。

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.