Third Harmonic Generation microscopy as a reliable diagnostic tool for evaluating lipid body modification during cell activation: The example of BV-2 microglia cells

Gavgiotaki, Evangelia; Filippidis, George; Kalognomou, M.; Tsouko, A.A.; Skordos, Ioannis; Fotakis, C.; Athanassakis, Irene

doi:10.1016/j.jsb.2014.11.011

Cited by 23 publications

(35 citation statements)

References 54 publications

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“…The experimental apparatus has been described analytically elsewhere [2] [4]. In brief, it consists of a custom made upright nonlinear microscope.…”

Section: Methodsmentioning

confidence: 99%

“…Image J program is employed for viewing and processing the obtained data. The developed system provides the capability to combine three nonlinear image modes in a single instrument, so that complementary information is taken regarding the structure and function of biological samples [2] [4]. To achieve polarized second and third harmonic generation (PSHG-PTHG) measurements an achromatic λ/2 wave plate (Thorlabs, AHWP05M-980) was added before the galvo silver mirrors to control and rotate the orientation of the excitation linear polarization.…”

Section: Methodsmentioning

confidence: 99%

“…The mean total area was calculated by setting a constant intensity threshold of THG signals that corresponds mainly in LBs [4]. Specifically the results shown in BV-2 cells that the mean total area , which corresponds mainly to LBs, was almost ten times bigger when the cells were activated (p<0.05) (Figure 3a).…”

Section: Depicts a Z Projection With Maximum Intensity Thg Image Of Bmentioning

confidence: 95%

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

et al. 2015

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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) andTwo 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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“…The experimental apparatus has been described analytically elsewhere [2] [4]. In brief, it consists of a custom made upright nonlinear microscope.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Depicts a Z Projection With Maximum Intensity Thg Image Of Bmentioning

confidence: 95%

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

et al. 2015

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“…Multi-photon microscopy (MPM) has been recognized as a promising technique for label-free imaging of biological samples [1,2]. Various nonlinear imaging modalities such as third harmonic generation (THG), second harmonic generation (SHG), two and three photon excited auto-fluorescence (2PEF/3PEF) can provide valuable information about intrinsic properties of the biological samples which can be employed in diagnosis applications [3][4][5][6]. However, the key challenge to take advantage of these imaging modalities in gastrointestinal and colonic visualization is to miniaturize the MPM into a compact and lightweight distal probe while keeping the general performance.…”

Section: Introductionmentioning

confidence: 99%

“…To the best of our knowledge, this is the first flexible endoscope which has been successfully employed for THG imaging. Recently, THG microscopy has been used by multiple groups for label-free pathology, optical biopsy and detecting cancerous cells [3][4][5][6]. Therefore, development of a flexible endoscope with THG imaging capability will pave the way for future research in this field.…”

Section: Introductionmentioning

confidence: 99%

Compact fiber-based multi-photon endoscope working at 1700 nm

Akhoundi

Qin

Peyghambarian

et al. 2018

Biomed. Opt. Express

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Abstract:We present the design, implementation and performance analysis of a compact multi-photon endoscope based on a piezo electric scanning tube. A miniature objective lens with a long working distance and a high numerical aperture (≈ 0.5) is designed to provide a diffraction limited spot size. Furthermore, a 1700 nm wavelength femtosecond fiber laser is used as an excitation source to overcome the scattering of biological tissues and reduce water absorption. Therefore, the novel optical system along with the unique wavelength allows us to increase the imaging depth. We demonstrate that the endoscope is capable of performing third and second harmonic generation (THG/SHG) and three-photon excitation fluorescence (3PEF) imaging over a large field of view (> 400 µm) with high lateral resolution (2.2 µm). The compact and lightweight probe design makes it suitable for minimally-invasive in-vivo imaging as a potential alternative to surgical biopsies.

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Distinction between breast cancer cell subtypes using third harmonic generation microscopy

Gavgiotaki

Filippidis

Markomanolaki

et al. 2016

Journal of Biophotonics

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Third Harmonic Generation (THG) microscopy as a non-invasive, label free imaging methodology, allows linkage of lipid profiles with various breast cancer cells. The collected THG signal arise mostly from the lipid droplets and the membrane lipid bilayer. Quantification of THG signal can accurately distinguish HER2-positive cells. Further analysis using Fourier transform infrared (FTIR) spectra reveals cancer-specific profiles, correlating lipid raft-corresponding spectra to THG signal, associating thus THG to chemical information. THG imaging of a cancer cell.

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Third Harmonic Generation microscopy as a reliable diagnostic tool for evaluating lipid body modification during cell activation: The example of BV-2 microglia cells

Cited by 23 publications

References 54 publications

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

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

Compact fiber-based multi-photon endoscope working at 1700 nm

Distinction between breast cancer cell subtypes using third harmonic generation microscopy

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