Ultrashort laser pulse cell manipulation using nano-and micro-materials

verfasst von: Markus Schomaker, Doreen Killian, Saskia Willenbrock, Eric Diebold, Eric Mazur, Willem Bintig, Anaclet Ngezahayo, Ingo Nolte, Hugo Murua Escobar, Christian Junghanß, Holger Lubatschowski, Alexander Heisterkamp
Abstract: The delivery of extra cellular molecules into cells is essential for cell manipulation. For this purpose genetic materials (DNA/RNA) or proteins have to overcome the impermeable cell membrane. To increase the delivery efficiency and cell viability of common methods different nano-and micro material based approaches were applied. To manipulate the cells, the membrane is in contact with the biocompatible material. Due to a field enhancement of the laser light at the material and the resulting effect the cell membrane gets perforated and extracellular molecules can diffuse into the cytoplasm. Membrane impermeable dyes, fluorescent labelled siRNA, as well as plasmid vectors encoded for GFP expression were used as an indicator for successful perforation or transfection, respectively. Dependent on the used material, perforation efficiencies over 90 % with a cell viability of about 80 % can be achieved. Additionally, we observed similar efficiencies for siRNA transfection. Due to the larger molecule size and the essential transport of the DNA into the nucleus cells are more difficult to transfect with GFP plasmid vectors. Proof of principle experiments show promising and adequate efficiencies by applying micro materials for plasmid vector transfection. For all methods a weakly focused fs laser beam is used to enable a high manipulation throughput for adherent and suspension cells. Furthermore, with these alternative optical manipulation methods it is possible to perforate the membrane of sensitive cell types such as primary and stem cells with a high viability.
Organisationseinheit(en): Institut für Zellbiologie und Biophysik
Externe Organisation(en): Laser Zentrum Hannover e.V. (LZH)
Universität Rostock
Stiftung Tierärztliche Hochschule Hannover
Harvard University
REBIRTH Forschungszentrum für translationale regenerative Medizin
Typ: Aufsatz in Konferenzband
Publikationsdatum: 27.08.2010
Publikationsstatus: Veröffentlicht
Peer-reviewed: Ja
ASJC Scopus Sachgebiete: Elektronische, optische und magnetische Materialien, Physik der kondensierten Materie, Angewandte Informatik, Angewandte Mathematik, Elektrotechnik und Elektronik
Elektronische Version(en): https://doi.org/10.1117/12.869054 (Zugang: Unbekannt)

BibTeX

@inproceedings{6c972667916e40c496d7069b4b25b942,
title = "Ultrashort laser pulse cell manipulation using nano-and micro-materials",
abstract = "The delivery of extra cellular molecules into cells is essential for cell manipulation. For this purpose genetic materials (DNA/RNA) or proteins have to overcome the impermeable cell membrane. To increase the delivery efficiency and cell viability of common methods different nano-and micro material based approaches were applied. To manipulate the cells, the membrane is in contact with the biocompatible material. Due to a field enhancement of the laser light at the material and the resulting effect the cell membrane gets perforated and extracellular molecules can diffuse into the cytoplasm. Membrane impermeable dyes, fluorescent labelled siRNA, as well as plasmid vectors encoded for GFP expression were used as an indicator for successful perforation or transfection, respectively. Dependent on the used material, perforation efficiencies over 90 % with a cell viability of about 80 % can be achieved. Additionally, we observed similar efficiencies for siRNA transfection. Due to the larger molecule size and the essential transport of the DNA into the nucleus cells are more difficult to transfect with GFP plasmid vectors. Proof of principle experiments show promising and adequate efficiencies by applying micro materials for plasmid vector transfection. For all methods a weakly focused fs laser beam is used to enable a high manipulation throughput for adherent and suspension cells. Furthermore, with these alternative optical manipulation methods it is possible to perforate the membrane of sensitive cell types such as primary and stem cells with a high viability.",
keywords = "Biophotonics, Cell manipulation, GFP, Membrane permeabilization, Nanomaterial, Nanoparticles, Perforation, Plasmonics, Transfection, Ultrashort laser pulses",
author = "Markus Schomaker and Doreen Killian and Saskia Willenbrock and Eric Diebold and Eric Mazur and Willem Bintig and Anaclet Ngezahayo and Ingo Nolte and {Murua Escobar}, Hugo and Christian Junghan{\ss} and Holger Lubatschowski and Alexander Heisterkamp",
year = "2010",
month = aug,
day = "27",
doi = "10.1117/12.869054",
language = "English",
isbn = "9780819482587",
series = "Proceedings of SPIE - The International Society for Optical Engineering",
booktitle = "Optical Trapping and Optical Micromanipulation VII",
note = "Optical Trapping and Optical Micromanipulation VII ; Conference date: 01-08-2010 Through 05-08-2010",
}