title: Optoinjection for efficient targeted delivery of a broad range of compounds and macromolecules into diverse cell types.
abstract: 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.
authors.affiliation: Cyntellect, Inc., 6620 Mesa Ridge Road, Suite 100, San Diego, California 92121-2906, USA.
authors.forename: Imran B
authors.initials: IB
authors.lastname: Clark
authors.forename: Elie G
authors.initials: EG
authors.lastname: Hanania
authors.forename: Janine
authors.initials: J
authors.lastname: Stevens
authors.forename: Marijo
authors.initials: M
authors.lastname: Gallina
authors.forename: Annabeth
authors.initials: A
authors.lastname: Fieck
authors.forename: Rolf
authors.initials: R
authors.lastname: Brandes
authors.forename: Bernhard O
authors.initials: BO
authors.lastname: Palsson
authors.forename: Manfred R
authors.initials: MR
authors.lastname: Koller
mesh_terms: D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism* / Q000528:radiation effects*; D002463:Cell Membrane Permeability / Q000528:radiation effects*; D006224:Cricetinae; D004307:Dose-Response Relationship, Radiation; D016503:Drug Delivery Systems / Q000379:methods*; D006801:Humans; D007834:Lasers; D051379:Mice; D008845:Microinjections / Q000379:methods*; D004364:Pharmaceutical Preparations / Q000008:administration & dosage*; D010599:Pharmacokinetics; D011829:Radiation Dosage; D013045:Species Specificity; D013314:Stress, Mechanical
chemical_list: D004364:Pharmaceutical Preparations