Estimation of the effective orientation of the SHG source in primary cortical neurons

Psilodimitrakopoulos, Sotiris; Petegnief, Valérie; Sòria, Guadalupe; Amat-Roldán, Ivan; Artigas, David; Planas, Anna M.; Loza-Álvarez, Pablo

doi:10.1364/oe.17.014418

Cited by 40 publications

(43 citation statements)

References 29 publications

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“…Note that a frequency-doubled version of the same fiber laser source could be used for a multimodal microscopy approach, allowing the implementation of other techniques such as TPEF, 35 SHG, 14,15,[19][20][21][22]25 etc. Adding these techniques will comprise an improved and supporting tool that would provide complementary information useful to investigate the source of the THG signal.…”

Section: Resultsmentioning

confidence: 99%

“…These valuable tools have been used to study in vivo biological samples ͑i.e., C. elegans, Drosophila melanogaster, and Zebrafish͒. [14][15][16][17][18][19][20][21][22] For instance, these studies aim to establish higher harmonic imaging modalities as label-free techniques showing a great potential to perform different developmental studies in a less invasive way. The generation of SHG signal requires a noncentrosymmetric structure, limiting its applicability in tissue mainly to collagen, muscle, and microtubules.…”

Section: Introductionmentioning

confidence: 99%

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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: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Third-harmonic generation for the study of Caenorhabditis elegans embryogenesis

et al. 2010

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“…The Fourier coefficients [Eq. (6)] are then to be replaced by E Q -T A R G E T ; t e m p : i n t r a l i n k -; e 0 0 9 ; 6 3 ; 5 4 4 a 0…”

Section: Discussionmentioning

confidence: 99%

“…The well-defined properties of the SHG signal makes it convenient to study well-ordered structures, such as collagen type I rich structures, 1-4 microtubules inside mitotic spindles or neurons [5][6][7] and striated muscle containing myosin thick filaments. [8][9][10][11] SHG microscopy relies on the presence of noncentrosymmetric features, achieved through a high degree of order of polar biomolecules.…”

Section: Introductionmentioning

confidence: 99%

On the interpretation of second harmonic generation intensity profiles of striated muscle

et al. 2015

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Abstract. Recently, a supramolecular model was developed for predicting striated skeletal muscle intensity profiles obtained by label-free second harmonic generation (SHG) microscopy. This model allows for a quantitative determination of the length of the thick filament antiparallel range or M-band (M), and results in M ¼ 0.12 μm for single-band intensity profiles when fixing the A-band length (A) to A ¼ 1.6 μm, a value originating from electron microscopy (EM) observations. Using simulations and experimental data sets, we showed that the objective numerical aperture (NA) and the refractive index (RI) mismatch (Δn ¼ n 2ω − n ω ) between the illumination wave (ω) and the second harmonic wave (2ω) severely affect the simulated sarcomere intensity profiles. Therefore, our recovered filament lengths did not match with those observed by EM. For an RI mismatch of Δn ¼ 0.02 and a moderate illumination NA of 0.8, analysis of single-band SHG intensity profiles with freely adjustable A-and M-band sizes yielded A ¼ 1.40 AE 0.04 μm and M ¼ 0.07 AE 0.05 μm for skeletal muscle. These lower than expected values were rationalized in terms of the myosin density distribution along the myosin thick filament axis. Our data provided new and practical insights for the application of the supramolecular model to study SHG intensity profiles in striated muscle.

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“…In this figure, some parts of the neurons forming the nerve ring are also observed even though a narrow-band SHG filter was used. This could be possibly due to fluorescence leaking through the SHG filter or SHG emission from the neuron [61]. Mapping the TPEF and SHG signals ( Figure 5.12c) enables the observation of how the neurons forming the nerve ring innervate with the isthmus of the pharynx.…”

Section: Nonlinear Imaging Testsmentioning

confidence: 99%