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.
|Zordan, Michael D; Mill, Christopher P; Riese 2nd, David J et al. (2011) A high throughput, interactive imaging, bright-field wound healing assay. Cytometry A 79:227-32|