title: High-throughput continuous flow femtosecond laser-assisted cell optoporation and transfection.
abstract: We present a femtosecond-laser based nanoprocessing system for transient optical cell membrane poration to allow the introduction of foreign molecules into the interior of a cell with very high throughput. In the setup, cells flow through a micro-flow tube for spatial confinement and are simultaneously targeted by fs laser radiation. Beam-shaping generates a focal geometry along a line which is scanned across the micro-flow cell to increase the number of reachable cells. Successful cell membrane poration was observed indirectly by cell transfection even with cell-light interaction times in the millisecond range. The system was characterized by experiments with Chinese hamster ovary cells regarding cell viability, the uptake of extrinsic molecules and cell transfection efficiency. The continuous flow of cells enables a tremendous increase of cell throughput compared to previous nonflow approaches by treating millions of cells, although with only limited efficiency. The setup opens the possibility to realize a completely automated high-throughput laser-assisted cell-poration system which could be integrated in lab-on-a-chip devices.
authors.affiliation: Department of Biophotonics and Laser Technology, Saarland University, Faculty of Mechatronics and Physics, Campus A5.1, 66123, Saarbrücken, Germany; JenLab GmbH, Schillerstr. 1, 07745 Jena, Germany and Science Park 2, Campus D1.2, 66123, Saarbrücken, Germany.
authors.forename: Hans Georg
authors.initials: HG
authors.lastname: Breunig
authors.forename: Aisada
authors.initials: A
authors.lastname: Uchugonova
authors.forename: Ana
authors.initials: A
authors.lastname: Batista
authors.forename: Karsten
authors.initials: K
authors.lastname: König
mesh_terms: D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000502:physiology / Q000528:radiation effects; D002463:Cell Membrane Permeability / Q000528:radiation effects; D006224:Cricetinae; D003412:Cricetulus; D005434:Flow Cytometry / Q000379:methods; D007834:Lasers; D014162:Transfection / Q000379:methods*
keywords: Bessel beam; cell transfection; flow cell; laser-assisted poration; optical transfection; optoporation