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

Imran B. Clark, Elie G. Hanania, Janine Stevens, Marijo Gallina, Annabeth Fieck, Rolf Brandes, Bernhard O. Palsson, Manfred R. Koller*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

56 Citations (Scopus)

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.

Original languageEnglish
Article number014034
JournalJournal of Biomedical Optics
Volume11
Issue number1
DOIs
Publication statusPublished - Jan 2006

Bibliographical note

Funding Information:
This material is based in part on work supported by the National Science Foundation (NSF) under Award No. DMI-0321740 and by the National Institutes of Health (NIH) under Award Nos. R44RR15374 and R44HG002985. Any opinions, findings, and conclusions or recommendations expressed in this publication are those of the authors and do not necessarily reflect the views of NSF or NIH. The authors are employees or direct affiliates of Cyntellect, Inc.

Other keywords

  • Cell transfection
  • Ions
  • Laser
  • Proteins
  • Semiconductor nanocrystals
  • Small interfering RNA

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