title: Characterization of the cellular response triggered by gold nanoparticle-mediated laser manipulation.
abstract: Laser-based transfection techniques have proven high applicability in several cell biologic applications. The delivery of different molecules using these techniques has been extensively investigated. In particular, new high-throughput approaches such as gold nanoparticle–mediated laser transfection allow efficient delivery of antisense molecules or proteins into cells preserving high cell viabilities. However, the cellular response to the perforation procedure is not well understood. We herein analyzed the perforation kinetics of single cells during resonant gold nanoparticle–mediated laser manipulation with an 850-ps laser system at a wavelength of 532 nm. Inflow velocity of propidium iodide into manipulated cells reached a maximum within a few seconds. Experiments based on the inflow of FM4-64 indicated that the membrane remains permeable for a few minutes for small molecules. To further characterize the cellular response postmanipulation, we analyzed levels of oxidative heat or general stress. Although we observed an increased formation of reactive oxygen species by an increase of dichlorofluorescein fluorescence, heat shock protein 70 was not upregulated in laser-treated cells. Additionally, no evidence of stress granule formation was visible by immunofluorescence staining. The data provided in this study help to identify the cellular reactions to gold nanoparticle–mediated laser manipulation.
authors.affiliation: Laser Zentrum Hannover e.V., Department of Biomedical Optics, Hollerithallee 8, Hannover 30419, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Kalies
authors.affiliation: Laser Zentrum Hannover e.V., Department of Biomedical Optics, Hollerithallee 8, Hannover 30419, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Keil
authors.affiliation: Laser Zentrum Hannover e.V., Department of Biomedical Optics, Hollerithallee 8, Hannover 30419, Germany.
authors.forename: Sina
authors.initials: S
authors.lastname: Sender
authors.affiliation: University of Veterinary Medicine, Small Animal Clinic, Bünteweg 9, Hannover 30599, GermanycUniversity of Rostock, Department of Hematology, Oncology, and Palliative Medicine, Ernst-Heydemann-Street 6, Rostock 18057, Germany.
authors.forename: Susanne C
authors.initials: SC
authors.lastname: Hammer
authors.affiliation: Laser Zentrum Hannover e.V., Department of Biomedical Optics, Hollerithallee 8, Hannover 30419, Germany.
authors.forename: Georgios C
authors.initials: GC
authors.lastname: Antonopoulos
authors.affiliation: Laser Zentrum Hannover e.V., Department of Biomedical Optics, Hollerithallee 8, Hannover 30419, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Schomaker
authors.affiliation: Laser Zentrum Hannover e.V., Department of Biomedical Optics, Hollerithallee 8, Hannover 30419, Germany.
authors.forename: Tammo
authors.initials: T
authors.lastname: Ripken
authors.affiliation: University of Veterinary Medicine, Small Animal Clinic, Bünteweg 9, Hannover 30599, GermanycUniversity of Rostock, Department of Hematology, Oncology, and Palliative Medicine, Ernst-Heydemann-Street 6, Rostock 18057, Germany.
authors.forename: Hugo
authors.initials: H
authors.lastname: Murua Escobar
authors.affiliation: Laser Zentrum Hannover e.V., Department of Biomedical Optics, Hollerithallee 8, Hannover 30419, Germany.
authors.forename: Heiko
authors.initials: H
authors.lastname: Meyer
authors.affiliation: Laser Zentrum Hannover e.V., Department of Biomedical Optics, Hollerithallee 8, Hannover 30419, Germany.
authors.forename: Dag
authors.initials: D
authors.lastname: Heinemann
mesh_terms: D002463:Cell Membrane Permeability / Q000502:physiology* / Q000528:radiation effects; D018274:Electroporation / Q000379:methods*; D006046:Gold / Q000737:chemistry / Q000528:radiation effects*; D053768:Metal Nanoparticles / Q000737:chemistry / Q000528:radiation effects*; D052898:Optical Tweezers; D011829:Radiation Dosage; D014162:Transfection / Q000379:methods*
chemical_list: D006046:Gold