Imaging the bipolarity of myosin filaments with Interferometric Second Harmonic Generation microscopy

Rivard, Maxime; Couture, Charles André; Miri, Amir K.; Laliberté, Mathieu; Bertrand-Grenier, Antony; Mongeau, Luc; Légaré, François

doi:10.1364/boe.4.002078

Cited by 22 publications

(18 citation statements)

References 34 publications

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“…Specifically, SHG microscopy has been used to image connective tissues rich in fibrillar collagen such as tendon [16,17], cornea [18,19], skin [20,21], fascia [22,23], cartilage [24,25]. In addition, SHG signals have been obtained from the myosin band in skeletal muscles [14,26] and from tubulin forming the microtubules in cultures of neurons [27,28] or during cell mitosis [1,29]. The common property of these three proteins (fibrillar collagen, myosin and tubulin) is their non-centrosymmetric structure at the macro-molecular scale.…”

Section: Introductionmentioning

confidence: 99%

“…Recently, its potential for tissue imaging has been demonstrated. In particular, I-SHG has been used to investigate the bipolarity of myosin filaments in skeletal muscles [26] and the polarity switch in collagen fascicles in tendon [34]. Note that only few other techniques allow to probe the relative polarity in tissues such as holographic SHG [35,36] or interferometric sum-frequency generation (SFG) [37].…”

Section: Introductionmentioning

confidence: 99%

“…However, in its first implementation, I-SHG suffered from two important drawbacks. First, the picosecond pulses excitation [26,34], used to simplify the interferometric part of the setup, decreased drastically the imaging contrast in tissues because of the smaller excitation peak intensity obtained. In practice, while this low contrast was enough to image highly organized structures such as tendon or muscles, it limited the investigation of more complex architectures such as skin or cartilage.…”

Section: Introductionmentioning

confidence: 99%

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Fast interferometric second harmonic generation microscopy

Bancelin¹,

Couture²,

Légaré³

et al. 2016

Biomed. Opt. Express

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We report the implementation of fast Interferometric Second Harmonic Generation (I-SHG) microscopy to study the polarity of non-centrosymmetric structures in biological tissues. Using a sample quartz plate, we calibrate the spatially varying phase shift introduced by the laser scanning system. Compensating this phase shift allows us to retrieve the correct phase distribution in periodically poled lithium niobate, used as a model sample. Finally, we used fast interferometric second harmonic generation microscopy to acquire phase images in tendon. Our results show that the method exposed here, using a laser scanning system, allows to recover the polarity of collagen fibrils, similarly to standard I-SHG (using a sample scanning system), but with an imaging time about 40 times shorter.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Fast interferometric second harmonic generation microscopy

Bancelin¹,

Couture²,

Légaré³

et al. 2016

Biomed. Opt. Express

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“…Although the mean value and the variance of noise intensity decreased significantly when the borosilicate substrate material was replaced with high-purity SiO 2 , the noise level persisted. This unexpected noise has not been considered in previous SHG studies of bulk samples 3,[5][6][7][8][9][10][11][12][13]30 . The problem might be negligible when the laser focus is significantly far from the substrate, or when the signal intensity generated by the specimen is sufficiently high.…”

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confidence: 99%

Second harmonic generation polarization microscopy as a tool for protein structure analysis

Kaneshiro

Okada

Shima

et al. 2018

Preprint

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Second-harmonic generation (SHG) is a nonlinear coherent scattering process that is sensitiveto molecular structures in illuminated materials. We report SHG polarization measurement for the detection of protein conformational changes in solutions of macromolecular protein assemblies such as microtubules and protein crystals. The results illustrate the potential of this method for protein structural analysis in physiological solutions at room temperature without labelling.Second-harmonic generation (SHG) microscopy is a powerful modality that enables the visualization of fibre structures without labelling. Such structures include the fibrillar collagen in tendons, myosin thick filaments in myosarcoma, and the densely bundled microtubules in living specimens 1-3 . SHG is a coherent process in which two photons that have the same energy are effectively 'combined' to generate a new photon with twice the energy; it was first discovered in 1961 4 . Twenty-five years later, SHG was used to determine the orientation of collagen fibres in a rat tail tendon 5 . Because SHG is sensitive to static electric field, SHG microscopy was also used to measure the action potential of a nerve cell 6,7 . The microscope design is based on that used for laser-scanning confocal fluorescence microscopes, so that the simultaneous observation of fluorescence and SHG is possible 8 .SHG is now also used for whole-animal imaging in vivo 9,10 .The nonlinear susceptibility of a material to SHG is expressed as the 3 rd -rank tensor (SHG tensor) originating from molecules with broken centrosymmetry and their alignments in macromolecular assemblies 1,2 . This unique feature may enable the acquisition of data about the spatial distributions of filament orientations, and can provide information on protein secondary structures, such as the axial pitch of alpha-helices 11 . Furthermore, SHG microscopy can be used to visualize the spatial distribution of fibre polarity directions in tissues with interferometric configurations 12,13 . Therefore, the SHG signal potentially carries structural information about proteins, which can be recorded optically without labelling.

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“…This feature issue is a compilation of papers by selected authors who presented their work at the NTM symposium and others active in the area of technique development. In this feature, new methods in nonlinear optical imaging are discussed [2][3][4][5], including approaches for deeper imaging [2] and interferometric nonlinear optical techniques to retrieve information about molecular orientation [4]. Exciting new developments in linear interferometric techniques are also highlighted [6-9], among which clever implementations of spatial light modulators for achieving better contrast [8,9].…”

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confidence: 99%