In vivo imaging of cell morphology and cellular processes in Caenorhabditis elegans, using non-linear phenomena

Filippidis, George; Gualda, Emilio J.; Mari, Meropi; Troulinaki, Kostoula; Fotakis, C.; Tavernarakis, Nektarios

doi:10.1016/j.micron.2009.06.005

Cited by 17 publications

(11 citation statements)

References 14 publications

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“…27 Given this, one might think that this wavelength cannot be used for highresolution imaging of living samples, nor for long-term imaging experiments. However, in our case, and in agreement with other works, [16][17][18][19]21,[24][25][26]34 the laser used is able to produce an efficient THG signal from the sample using very low average powers. All this helps reduce the expected damage.…”

Section: Resultssupporting

confidence: 92%

Third-harmonic generation for the study of Caenorhabditis elegans embryogenesis

et al. 2010

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Abstract. Live microscopy techniques ͑i.e., differential interference contrast, confocal microscopy, etc.͒ have enabled the understanding of the mechanisms involved in cells and tissue formation. In long-term studies, special care must be taken in order to avoid sample damage, restricting the applicability of the different microscopy techniques. We demonstrate the potential of using third-harmonic generation ͑THG͒ microscopy for morphogenesis/embryogenesis studies in living Caenorhabditis elegans ͑C. elegans͒. Moreover, we show that the THG signal is obtained in all the embryo development stages, showing different tissue/structure information. For this research, we employ a 1550-nm femtosecond fiber laser and demonstrate that the expected water absorption at this wavelength does not severely compromise sample viability. Additionally, this has the important advantage that the THG signal is emitted at visible wavelengths ͑516 nm͒. Therefore, standard collection optics and detectors operating near maximum efficiency enable an optimal signal reconstruction. All this, to the best of our knowledge, demonstrates for the first time the noninvasiveness and strong potential of this particular wavelength to be used for highresolution four-dimensional imaging of embryogenesis using unstained C. elegans in vivo samples. © 2010 Society of Photo-Optical Instrumentation Engineers.

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Section: Resultssupporting

confidence: 92%

Third-harmonic generation for the study of Caenorhabditis elegans embryogenesis

et al. 2010

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“…THG is a powerful noninvasive imaging technique capable of revealing the anatomic features of many model organisms such as C. elegans in vivo (Gualda et al, 2008). THG requires a single beam and has already proven its efficiency to monitor cellular processes in vivo (Filippidis et al, 2009), but also in vitro combined with other nonlinear modalities (Aptel et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Imaging ectopic fat deposition incaenorhabditis elegansmuscles using nonlinear microscopy

Mari

Filippidis

Palikaras

et al. 2015

Microsc. Res. Tech.

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The elucidation of the molecular mechanisms that lead to the development of metabolic syndrome, a complex of pathological conditions including type-2 diabetes, hypertension, and cardiovascular diseases, is an important issue with high biological significance and requires accurate methods capable of monitoring lipid storage distribution and dynamics in vivo. In this study, the nonlinear phenomena of second and third harmonic generation (SHG, THG) have been employed simultaneously as label-free, nondestructive diagnostic techniques, for the monitoring and the complementary three-dimensional (3D) imaging and analysis of the muscular areas and the lipid content localization. THG microscopy was used as a quantitative tool in order to record the accumulation of lipids in nonadipose tissues in the pharyngeal muscles of 18 Caenorhabditis elegans (C. elegans) specimens, while the SHG imaging provided the detailed anatomical information about the structure of the muscles. The ectopic accumulation of fat on the pharyngeal muscles increases in wild-type (N2) C. elegans between 1 and 9 days of adulthood. This suggests a correlation of ectopic fat accumulation with the process of aging. Our results can contribute to the unraveling of the link between the deposition of ectopic fat and aging, but mainly to the validation of SHG and THG microscopy modalities as new, noninvasive tools to localize and quantify selectively lipid formation and distribution.

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“…THG is a powerful non-invasive imaging technique able to reveal the anatomic features of many model organisms among them is the C. elegans 15 . THG requires a single beam and is efficient of recording cellular processes in vivo 16 , but also in vitro combined with further non-linear modalities 17 .…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2015

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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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In vivo imaging of cell morphology and cellular processes in Caenorhabditis elegans, using non-linear phenomena

Cited by 17 publications

References 14 publications

Third-harmonic generation for the study of Caenorhabditis elegans embryogenesis

Third-harmonic generation for the study of Caenorhabditis elegans embryogenesis

Imaging ectopic fat deposition incaenorhabditis elegansmuscles using nonlinear microscopy

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

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