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Research Papers

Optoinjection for efficient targeted delivery of a broad range of compounds and macromolecules into diverse cell types

[+] Author Affiliations
Imran B. Clark, Elie G. Hanania, Janine Stevens, Marijo Gallina, Annabeth Fieck, Rolf Brandes, Bernhard O. Palsson, Manfred R. Koller

Cyntellect, Inc., 6620 Mesa Ridge Road, Suite 100, San Diego, California 92121–2906

J. Biomed. Opt. 11(1), 014034 (February 06, 2006). doi:10.1117/1.2168148
History: Received March 18, 2005; Revised August 19, 2005; Accepted September 13, 2005; Published February 06, 2006
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Efficient delivery of compounds and macromolecules into living cells is essential in many fields including basic research, applied drug discovery, and clinical gene therapy. Unfortunately, current delivery methods, such as cationic lipids and electroporation, are limited by the types of macromolecules and cells that can be employed, poor efficiency, and/or cell toxicity. To address these issues, novel methods were developed based on laser-mediated delivery of macromolecules into cells through optoinjection. An automated high-throughput instrument, the laser-enabled analysis and processing (LEAP™) system, was utilized to elucidate and optimize several parameters that influence optoinjection efficiency and toxicity. Techniques employing direct cell irradiation (i.e., targeted to specific cell coordinates) and grid-based irradiation (i.e., without locating individual cells) were both successfully developed. With both techniques, it was determined that multiple, sequential low radiant exposures produced more favorable results than a single high radiant exposure. Various substances were efficiently optoinjected—including ions, small molecules, dextrans, siRNAs (small interfering RNAs), plasmids, proteins, and semiconductor nanocrystals—into numerous cell types. Notably, cells refractory to traditional delivery methods were efficiently optoinjected with lower toxicity. We establish the broad utility of optoinjection, and furthermore, are the first to demonstrate its implementation in an automated, high-throughput manner.

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© 2006 Society of Photo-Optical Instrumentation Engineers

Citation

Imran B. Clark ; Elie G. Hanania ; Janine Stevens ; Marijo Gallina ; Annabeth Fieck, et al.
"Optoinjection for efficient targeted delivery of a broad range of compounds and macromolecules into diverse cell types", J. Biomed. Opt. 11(1), 014034 (February 06, 2006). ; http://dx.doi.org/10.1117/1.2168148


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