CAREER: Optics in Biological Research, Education, and Outreach

Cornell University, Ithaca NY

Investigators

Abstract

0846534 Schaffer "This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5)." The PI proposes to develop tools to enable the introduction of exogenous DNA into specifically targeted cells in the brain of a live rodent, opening the door to cell-targeted genetic manipulation. This new capability will extend the power of genetic manipulation techniques to a new class of experiments, where single-cell targeting enables cell-cell interactions and early stages of disease to be studied, in vivo. They will use tightly-focused femtosecond laser pulses to ablate a small, transient hole in the membrane of a targeted cell, allowing DNA that has been injected into the extracellular space to enter the cell. This hole rapidly reseals, leaving the cell alive and functional. Preliminary data confirms optoporation of targeted cells in vitro and in vivo as well as in vitro expression of optically transfected DNA. They will extend this technique to in vivo transfection in targeted brain cells. They further propose to use this technology to express light-gated ion channels in single brain neurons and use cell-resolved calcium imaging to follow how the optically initiated firing of the transfected cell effects activity in surrounding cells. This work could provide a unique means to study the functional connectivity of cortical neuronal circuits, in vivo.