title: Femtosecond laser nanosurgery of sub-cellular structures in HeLa cells by employing Third Harmonic Generation imaging modality as diagnostic tool.
abstract: Femtosecond laser assisted nanosurgery of microscopic biological specimens is a relatively new technique which allows the selective disruption of sub-cellular structures without causing any undesirable damage to the surrounding regions. The targeted structures have to be stained in order to be clearly visualized for the nanosurgery procedure. However, the validation of the final nanosurgery result is difficult, since the targeted structure could be simply photobleached rather than selectively destroyed. This fact comprises a main drawback of this technique. In our study we employed a multimodal system which integrates non-linear imaging modalities with nanosurgery capabilities, for the selective disruption of sub-cellular structures in HeLa cancer cells. Third Harmonic Generation (THG) imaging modality was used as a tool for the identification of structures that were subjected to nanosurgery experiments. No staining of the biological samples was required, since THG is an intrinsic property of matter. Furthermore, cells' viability after nanosurgery processing was verified via Two Photon Excitation Fluorescence (TPEF) measurements.
authors.affiliation: Institute of Electronic Structure and Laser, Foundation of Research and Technology-Hellas, P.O. Box 1385, 71110, Heraklion, Crete, Greece.
authors.forename: George J
authors.initials: GJ
authors.lastname: Tserevelakis
authors.forename: Stylianos
authors.initials: S
authors.lastname: Psycharakis
authors.forename: Bojan
authors.initials: B
authors.lastname: Resan
authors.forename: Felix
authors.initials: F
authors.lastname: Brunner
authors.forename: Evagelia
authors.initials: E
authors.lastname: Gavgiotaki
authors.forename: Kurt
authors.initials: K
authors.lastname: Weingarten
authors.forename: George
authors.initials: G
authors.lastname: Filippidis
mesh_terms: D002470:Cell Survival; D004867:Equipment Design; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods; D021621:Imaging, Three-Dimensional; D007834:Lasers; D008027:Light; D008055:Lipids / Q000737:chemistry; D008853:Microscopy / Q000379:methods; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D055095:Optics and Photonics
chemical_list: D008055:Lipids