title: Software-aided automatic laser optoporation and transfection of cells.
abstract: Optoporation, the permeabilization of a cell membrane by laser pulses, has emerged as a powerful non-invasive and highly efficient technique to induce transfection of cells. However, the usual tedious manual targeting of individual cells significantly limits the addressable cell number. To overcome this limitation, we present an experimental setup with custom-made software control, for computer-automated cell optoporation. The software evaluates the image contrast of cell contours, automatically designates cell locations for laser illumination, centres those locations in the laser focus, and executes the illumination. By software-controlled meandering of the sample stage, in principle all cells in a typical cell culture dish can be targeted without further user interaction. The automation allows for a significant increase in the number of treatable cells compared to a manual approach. For a laser illumination duration of 100 ms, 7-8 positions on different cells can be targeted every second inside the area of the microscope field of view. The experimental capabilities of the setup are illustrated in experiments with Chinese hamster ovary cells. Furthermore, the influence of laser power is discussed, with mention on post-treatment cell survival and optoporation-efficiency rates.
authors.affiliation: Saarland University, Faculty of Mechatronics and Physics, Department of Biophotonics and Laser Technology, 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
authors.initials: H
authors.lastname: Georg Breunig
authors.affiliation: Saarland University, Faculty of Mechatronics and Physics, Department of Biophotonics and Laser Technology, 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: Aisada
authors.initials: A
authors.lastname: Uchugonova
authors.affiliation: Saarland University, Faculty of Mechatronics and Physics, Department of Biophotonics and Laser Technology, Campus A5.1, 66123 Saarbrücken, Germany.
authors.forename: Ana
authors.initials: A
authors.lastname: Batista
authors.affiliation: Saarland University, Faculty of Mechatronics and Physics, Department of Biophotonics and Laser Technology, 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: Karsten
authors.initials: K
authors.lastname: König
mesh_terms: D000818:Animals; D057205:Automation, Laboratory / Q000379:methods*; D016466:CHO Cells; D018929:Cell Culture Techniques; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D003412:Cricetulus; D007834:Lasers; D014162:Transfection / Q000379:methods*