Targeted microinjection into cells and retina using optoporation

Gu, Ling; Mohanty, Samarendra K.

doi:doi:10.1117/1.3662887

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Journal of Biomedical Optics / Volume 16 / Issue 12 / Research Papers: Therapeutic

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Targeted microinjection into cells and retina using optoporation

J. Biomed. Opt. 16, 128003 (Dec 06, 2011); http://dx.doi.org/10.1117/1.3662887

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Abstract

References (19)

Ling Gu and Samarendra K. Mohanty

University of Texas at Arlington, Department of Physics, Biophysics and Physiology Lab, Arlington, Texas 76019

The laser microbeam has enabled highly precise noncontact delivery of exogenous materials into targeted cells without compromising cell viability, which has been a highly challenging task for traditional methods. Here, we report targeted delivery of impermeable substances into mammalian cells and goldfish retinal explants subsequent to ultrafast laser microbeam assisted injection. Introduction of impermeable dye into the cell through localized pore formation was confirmed by distinct fluorescence at the site of pore formation on the membrane and its spatiotemporal diffusion pattern through the nucleus. Indirect optoporation by bubble formation, external to cell, led to a similar spatial diffusion pattern but with a larger time constant for injection. Using optimized laser intensity, exposure, and a spatial irradiation pattern, desired spatial transfection patterns in goldfish retina explants were achieved as confirmed by the expression of injected plasmids encoded for light-activable channelrhodopsin-2 ion-channel, tagged with fluorescent protein. Laser assisted delivery of exogenous material into a specific area of three-dimensional neuronal tissue, such as the retina, will help to understand the functioning of neuronal circuitry of normal and degenerated retina.

History

Received May 24, 2011
Accepted Nov 01, 2011
Revised Oct 22, 2011
Published online Dec 06, 2011

Digital Object Identifier

http://dx.doi.org/10.1117/1.3662887

Citation

Ling Gu and Samarendra K. Mohanty, "Targeted microinjection into cells and retina using optoporation", J. Biomed. Opt. 16, 128003 (Dec 06, 2011); http://dx.doi.org/10.1117/1.3662887