Third Harmonic Generation microscopy as a diagnostic tool for the investigation of microglia BV-2 and breast cancer cells activation

E. Gavgiotaki; G. Filippidis; S. Psilodimitrakopoulos; H. Markomanolaki; M. Kalognomou; S. Agelaki; V. Georgoulias; I. Athanassakis

doi:10.1117/12.2183286

14 July 2015 Third Harmonic Generation microscopy as a diagnostic tool for the investigation of microglia BV-2 and breast cancer cells activation

E. Gavgiotaki, G. Filippidis, S. Psilodimitrakopoulos, H. Markomanolaki, M. Kalognomou, S. Agelaki, V. Georgoulias, I. Athanassakis

Author Affiliations +

Proceedings Volume 9536, Advanced Microscopy Techniques IV; and Neurophotonics II; 953614 (2015) https://doi.org/10.1117/12.2183286
Event: European Conferences on Biomedical Optics, 2015, Munich, Germany

Abstract

Nonlinear optical imaging techniques have created new opportunities of research in the biomedical field. Specifically, Third Harmonic Generation (THG) seems to be a suitable noninvasive imaging tool for the delineation and quantification of biological structures at the microscopic level. The aim of this study was to extract information as to the activation state of different cell types by using the THG imaging microscopy as a diagnostic tool.

BV-2 microglia cell line was used as a representative biological model enabling the study of resting and activated state of the cells linked to various pathological conditions. Third Harmonic Generation (THG) and Two Photon Excitation Fluorescence (TPEF) measurements were simultaneously collected from stained breast cancer cells, by employing a single homemade experimental apparatus and it was shown that high THG signals mostly arise from lipid bodies. Continuously, BV-2 microglia cells were examined with or without activation by lipopolysaccharide (LPS) in order to discriminate between control and activated cells based on the quantification of THG signals. Statistically quantification was accomplished in both mean area and mean intensity values of THG. The values for mean total area and mean THG intensity values have been increased in activated versus the non-activated cells. Similar studies of quantification are underway in breast cancer cells for the exact discrimination on different cell lines. Furthermore, laser polarization dependence of SHG and THG signal in unstained biological samples is investigated.

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E. Gavgiotaki, G. Filippidis, S. Psilodimitrakopoulos, H. Markomanolaki, M. Kalognomou, S. Agelaki, V. Georgoulias, and I. Athanassakis "Third Harmonic Generation microscopy as a diagnostic tool for the investigation of microglia BV-2 and breast cancer cells activation", Proc. SPIE 9536, Advanced Microscopy Techniques IV; and Neurophotonics II, 953614 (14 July 2015); https://doi.org/10.1117/12.2183286

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