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Femtosecond laser nanosurgery of sub‐cellular structures in HeLa cells by employing Third Harmonic Generation imaging modality as diagnostic tool
Author(s) -
Tserevelakis George J.,
Psycharakis Stylianos,
Resan Bojan,
Brunner Felix,
Gavgiotaki Evagelia,
Weingarten Kurt,
Filippidis George
Publication year - 2012
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201100055
Subject(s) - femtosecond , hela , laser , modality (human–computer interaction) , second harmonic generation , fluorescence , nanotechnology , biological imaging , materials science , two photon excitation microscopy , biological system , biophysics , optics , computer science , chemistry , cell , artificial intelligence , biology , physics , biochemistry
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. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)