Hannover 2016 – wissenschaftliches Programm
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MO: Fachverband Molekülphysik
MO 8: Experimental Techniques
MO 8.2: Vortrag
Dienstag, 1. März 2016, 11:15–11:30, f142
Optical cell transfection platform — •Hans Georg Breunig1, Ana Batista2, Aisada Uchugonova1,2, and Karsten König1,2 — 1JenLab GmbH, Science Park 2, 66123 Saarbrücken, Germany and Schillerstr. 1, 07745 Jena, Germany — 2Department of Biophotonics and Laser Technology, Saarland University, 66123 Saarbrücken, Germany
Cells can be transfected by transient laser-induced perforation of the cell membrane which allows foreign genetic material to enter the cell interior. This method (optoporation) has emerged as a powerful noninvasive and highly efficient cell-transfection technique. We present an experimental platform based on a modified multiphoton laser scanning microscope employing a femtosecond laser, beam shaping, and custom-made control software for computer-automated cell optoporation. The software evaluates the image contrast due to cell contours, automatically designates cell locations for laser illumination, centres those locations in the laser focus (Gaussian or Bessel beam profile), 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. For an illumination duration of 100 ms, 7-8 positions on different cells can be targeted every second. Ultra-short fs pulses are in particular efficient for the optoporation due to underlying multiphoton absorption processes. The experimental capabilities of the setup are illustrated in experiments with Chinese hamster ovary cells. Furthermore, the influence of laser characteristics on the optoporation efficiency is discussed.