Non-linear imaging techniques visualize the lipid profile of C. elegans

Meropi Mari; Barbara Petanidou; Konstantinos Palikaras; Costas Fotakis; Nektarios Tavernarakis; George Filippidis

doi:10.1117/12.2183161

14 July 2015 Non-linear imaging techniques visualize the lipid profile of C. elegans

Meropi Mari, Barbara Petanidou, Konstantinos Palikaras, Costas Fotakis, Nektarios Tavernarakis, George Filippidis

Proceedings Volume 9536, Advanced Microscopy Techniques IV; and Neurophotonics II; 953613 (2015) https://doi.org/10.1117/12.2183161
Event: European Conferences on Biomedical Optics, 2015, Munich, Germany

Abstract

The non-linear techniques Second and Third Harmonic Generation (SHG, THG) have been employed simultaneously to record three dimensional (3D) imaging and localize the lipid content of the muscular areas (ectopic fat) of Caenorhabditis elegans (C. elegans). Simultaneously, Two-Photon Fluorescence (TPEF) was used initially to localize the stained lipids with Nile Red, but also to confirm the THG potential to image lipids successfully. In addition, GFP labelling of the somatic muscles, proves the initial suggestion of the existence of ectopic fat on the muscles and provides complementary information to the SHG imaging of the pharynx. The ectopic fat may be related to a complex of pathological conditions including type-2 diabetes, hypertension and cardiovascular diseases. The elucidation of the molecular path leading to the development of metabolic syndrome is a vital issue with high biological significance and necessitates accurate methods competent of monitoring lipid storage distribution and dynamics in vivo. THG microscopy was employed as a quantitative tool to monitor the lipid accumulation in non-adipose tissues in the pharyngeal muscles of 12 unstained specimens while the SHG imaging revealed the anatomical structure of the muscles. The ectopic fat accumulation on the pharyngeal muscles increases in wild type (N2) C. elegans between 1 and 9 days of adulthood. This suggests a correlation of the ectopic fat accumulation with the aging. Our results can provide new evidence relating the deposition of ectopic fat with aging, but also validate SHG and THG microscopy modalities as new, non-invasive tools capable of localizing and quantifying selectively lipid accumulation and distribution.

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Meropi Mari, Barbara Petanidou, Konstantinos Palikaras, Costas Fotakis, Nektarios Tavernarakis, and George Filippidis "Non-linear imaging techniques visualize the lipid profile of C. elegans", Proc. SPIE 9536, Advanced Microscopy Techniques IV; and Neurophotonics II, 953613 (14 July 2015); https://doi.org/10.1117/12.2183161

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KEYWORDS

Second-harmonic generation

Head

Microscopy

Signal detection

Tissues

Green fluorescent protein

Visualization

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