Cellular Ion Control by Laser-Induced Permeabilization

Cyntellect, Inc., San Diego CA

Investigators

Abstract

DESCRIPTION (provided by applicant): Cells have intricate control mechanisms for regulating intracellular concentrations of several key ions, and ion imbalances often lead to dramatic consequences on cell physiology. Therefore, many researchers study how various ions and their concentrations affect cells, and the impact of various pharmaceutical agents on these parameters. Current techniques for altering cell ion levels include microinjection, compounds that form membrane channels (e.g., ionophores), release of intracellular calcium stores (by compounds or electrical stimulation) and electroporation. Recognized limitations with existing techniques provide an opportunity for an improved method which is rapid, reversible, broadly applicable (to many cell and ion types), and not damaging to cells. The Company has developed a novel instrument for high-throughput cell imaging and laser-based manipulation called LEAP. Preliminary results herein demonstrate utility of LEAP for laser-mediated optoinjection of ions into cells followed by kinetic imaging on an individual cell basis. The overall goal of this SBIR program is to develop technology for ion optoinjection and high-throughput kinetic imaging at the individual cell level as a new methodology for ion research, applicable to a number of important areas, including cardiac physiology, enzyme regulation, and cell death (apoptosis and necrosis). Specific Phase I aims include: (i) optimization of optoinjection parameters for maximum efficiency and viability, (ii) determining which cell types and ions are amenable to optoinjection, and (iii) initial characterization of the mechanism of ion optoinjection. Successful completion of Phase I will demonstrate that ion optoinjection can effectively alter intracellular concentrations of particular ions in experimentally important cell lines and primary cells. Further, use of the LEAP instrument platform for high-throughput kinetic imaging of many individual cells will be implemented and demonstrated. Phase II will then utilize this technology to study effects of altering the concentration of various ions on the physiological processes of cells and how various drugs interfere with such processes. Completion of Phase II will ultimately lead to the commercial development of novel tools and assays for basic ion physiology research and drug discovery.