title: Third Harmonic Generation microscopy as a reliable diagnostic tool for evaluating lipid body modification during cell activation: the example of BV-2 microglia cells.
abstract: Nonlinear optical processes have found widespread applications in fields ranging from fundamental physics to biomedicine. In this study, we attempted to evaluate cell activation by using the Third Harmonic Generation (THG) imaging microscopy as a new diagnostic tool. The BV-2 microglia cell line with or without activation by lipopolysaccharide was chosen as a representative biological model. The results showed that THG imaging could discriminate between the control versus activated state of BV-2 cells not only as to THG signal intensity but also as to THG signal area, while verifying once more that the majority of the intracellular detected signal corresponds to lipid bodies. Since THG imaging is a real time, non-destructive modality and does not require any prior cell processing and staining, the results presented here provide an important tool for normal versus activated cell discrimination, which could be proved very useful in the study of inflammation.
authors.affiliation: Institute of Electronic Structure and Laser, Foundation for Research and Technology Hellas, 71110 Heraklion, Greece; Department of Physics, University of Crete, Greece.
authors.forename: E
authors.initials: E
authors.lastname: Gavgiotaki
authors.affiliation: Institute of Electronic Structure and Laser, Foundation for Research and Technology Hellas, 71110 Heraklion, Greece.
authors.forename: G
authors.initials: G
authors.lastname: Filippidis
authors.affiliation: Department of Biology, University of Crete, Heraklion 71409, Crete, Greece.
authors.forename: M
authors.initials: M
authors.lastname: Kalognomou
authors.affiliation: Institute of Electronic Structure and Laser, Foundation for Research and Technology Hellas, 71110 Heraklion, Greece; Department of Physics, University of Crete, Greece.
authors.forename: A A
authors.initials: AA
authors.lastname: Tsouko
authors.affiliation: Department of Biology, University of Crete, Heraklion 71409, Crete, Greece.
authors.forename: I
authors.initials: I
authors.lastname: Skordos
authors.affiliation: Institute of Electronic Structure and Laser, Foundation for Research and Technology Hellas, 71110 Heraklion, Greece; Department of Physics, University of Crete, Greece.
authors.forename: C
authors.initials: C
authors.lastname: Fotakis
authors.affiliation: Department of Biology, University of Crete, Heraklion 71409, Crete, Greece. Electronic address: athan@biology.uoc.gr.
authors.forename: I
authors.initials: I
authors.lastname: Athanassakis
mesh_terms: D000818:Animals; D002460:Cell Line; D066292:Lipid Droplets / Q000648:ultrastructure*; D051379:Mice; D017628:Microglia / Q000502:physiology / Q000648:ultrastructure*; D036641:Microscopy, Fluorescence, Multiphoton
keywords: Activation; Imaging; Lipid bodies; Microglia cells; Nonlinear microscopy; Third Harmonic Generation