Multiplex coherent anti-Stokes Raman scattering highlights state of chromatin condensation in CH region

Ben, Tiffany Guerenne-Del; Rajaofara, Zakaniaina; Couderc, Vincent; Sol, Vincent; Kano, Hideaki; Leproux, Philippe; Petit, J.

doi:10.1038/s41598-019-50453-0

Cited by 28 publications

(29 citation statements)

References 42 publications

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“…These two spectral contributions slightly overlap, as in the case of colorectal cancer cell lines. The CH 2 signatures were mainly located close to the nucleus, and correspond to ER 15 , and their intensities increased in BDNF-treated cells . Signal intensity increased with treatment times and, after 72 h, a few lipid droplets appeared (Fig.…”

Section: Resultsmentioning

confidence: 97%

“…9 ). This signal includes the CARS signals of lipids, proteins and 5-methylcytosines of heterochromatin 15 . BODIPY staining revealed an accumulation of neutral lipids after 48 h of treatment, and a few lipid droplets appeared after 72 h, as shown with MCARS analysis (Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The spectroscopic setup used in this work is presented in 15 . The lateral, axial and spectral resolutions of the CARS microspectroscope were ~ 300 nm, 2 µm, and 0.8 cm −1 , respectively.…”

Section: Methodsmentioning

confidence: 99%

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Multiplex coherent anti-Stokes Raman scattering microspectroscopy detection of lipid droplets in cancer cells expressing TrkB

Ben

Couderc

Duponchel

et al. 2020

Sci Rep

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For many years, scientists have been looking for specific biomarkers associated with cancer cells for diagnosis purposes. These biomarkers mainly consist of proteins located at the cell surface (e.g. the TrkB receptor) whose activation is associated with specific metabolic modifications. Identification of these metabolic changes usually requires cell fixation and specific dye staining. MCARS microspectroscopy is a label-free, non-toxic, and minimally invasive method allowing to perform analyses of live cells and tissues. We used this method to follow the formation of lipid droplets in three colorectal cancer cell lines expressing TrkB. MCARS images of cells generated from signal integration of CH2 stretching modes allow to discriminate between lipid accumulation in the endoplasmic reticulum and the formation of cytoplasmic lipid droplets. We found that the number of the latter was related to the TrkB expression level. This result was confirmed thanks to the creation of a HEK cell line which over-expresses TrkB. We demonstrated that BDNF-induced TrkB activation leads to the formation of cytoplasmic lipid droplets, which can be abolished by K252a, an inhibitor of TrkB. So, MCARS microspectroscopy proved useful in characterizing cancer cells displaying an aberrant lipid metabolism.

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Section: Resultsmentioning

confidence: 97%

Section: Resultsmentioning

confidence: 99%

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Multiplex coherent anti-Stokes Raman scattering microspectroscopy detection of lipid droplets in cancer cells expressing TrkB

Ben

Couderc

Duponchel

et al. 2020

Sci Rep

Self Cite

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“…One direction is developing advanced high‐speed Raman microscopy systems, such as slit scanning Raman microscopy, [ 80 ] multifocal Raman microscopy, [ 80 ] direct Raman imaging systems, [ 81 ] and systems based on nonlinear optics. [ 82 ] Another direction is the use of plasmonic nanoparticles to enhance the Raman signals of biomolecules inside cells, [ 83 ] or outside cells, [ 84 ] which enables the change in the molecular signals related to the homeostasis of the cell to be monitored, as demonstrated in recent several papers. [ 83,85,86 ] However, high‐speed Raman imaging of a live single cell that does not sacrifice the image resolution and has the ability to track a single nanoparticle inside the cell in real time with spontaneous Raman scattering has not been firmly established.…”

Section: Bioanalytical Applicationsmentioning

confidence: 99%

Recent Advances in the Synthesis of Intra‐Nanogap Au Plasmonic Nanostructures for Bioanalytical Applications

Yang

Lim

2020

Advanced Materials

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frequency region (3100-3400 cm −1); therefore, the Raman spectrum of water does not interfere with the spectra of the molecules of interest, which is a critical property for bioanalytical applications. [3,4] However, the extremely low signal intensity of Raman scattering of molecules has been a major obstacle in the way of its wide application. [5] In contrast, fluorescence offers a high quantum yield in the visible region and generates high spatiotemporal images of the biological structures and functions upon appropriate modification with fluorescent dyes. The development of new optical techniques, such as confocal microscopy, total internal reflection fluorescence microscopy, and recently, super-resolution fluorescence microscopy, [6] has made it possible to obtain more detailed structural information. However, because fluorescence microscopy relies on labeling with external dye molecules, a fluorescence signal provides limited information and is prone to rapid photobleaching as major drawbacks for long-term monitoring. In the history of the development of Raman spectroscopy, two significant discoveries boosted research interest in the use of Raman scattering for practical applications. In 1973, Fleischmann et al. first reported the greatly enhanced Raman scattering intensity of pyridine molecules adsorbed on an electrochemically roughened silver surface, a technique known as surface-enhanced Raman scattering (SERS). [7] This effect was further explained by Jeanmaire and Van Duyne, who proposed an electromagnetic (EM) mechanism for the SERS. [8] The enhancement factor (EF) of the Raman signal intensity is proportional to the fourth order of the local EM field intensity (EF ∝|E| 4). [5] Albrecht and Creighton proposed that the chargetransfer effect contributes to SERS. [9] Both mechanisms are believed to be involved in enhancing the intensity of the Raman scattering spectrum. [10] Nearly 20 years later, in 1997, Nie and Kneipp independently suggested the possibility of single-molecule detection with Raman scattering from nanoparticle aggregates. [11,12] The EF values at this time were calculated to be in the range of 10 14-10 15 , which is believed to be the EF required to obtain a Raman scattering signal from a single molecule. [11,12] Zrimsek et al. and Park et al. experimentally demonstrated that an EF of ≈10 7-10 8 is sufficient to achieve single-molecule SERS. [10,13] Despite the controversy regarding the minimum EF for single molecule detection, it was clear that the presence of a Plasmonic nanogap-enhanced Raman scattering has attracted considerable attention in the fields of Raman-based bioanalytical applications and materials science. Various strategies have been proposed to prepare nanostructures with an inter-or intra-nanogap for fundamental study models or applications. This report focuses on recent advances in synthetic methods to fabricate intra-nanogap structures with diverse dimensions, with detailed focus on the theory and bioanalytical applications. Synthetic strategies ranging from the use of ...

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“…Published by Elsevier Inc. All rights reserved). (d) Analysis of interphase and mitotic, stained, fixed and living, HEK293 cells and fibroblasts (Adapted with permission from Guerenne‐Del Ben et al (2019) under the terms of the Creative Commons Attribution 4.0 International License http://creativecommons.org/licenses/by/4.0/)…”

Section: Typical Applications Of Coherent Raman Scattering Imaging Onmentioning

confidence: 99%

Mammalian cell and tissue imaging using Raman and coherent Raman microscopy

Fung

Shi

2020

WIREs Mechanisms of Disease

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Direct imaging of metabolism in cells or multicellular organisms is important for understanding many biological processes. Raman scattering (RS) microscopy, particularly, coherent Raman scattering (CRS) such as coherent anti-Stokes Raman scattering (CARS) and stimulated Raman scattering (SRS), has emerged as a powerful platform for cellular imaging due to its high chemical selectivity, sensitivity, and imaging speed. RS microscopy has been extensively used for the identification of subcellular structures, metabolic observation, and phenotypic characterization. Conjugating RS modalities with other techniques such as fluorescence or infrared (IR) spectroscopy, flow cytometry, and RNAsequencing can further extend the applications of RS imaging in microbiology, system biology, neurology, tumor biology and more. Here we overview RS modalities and techniques for mammalian cell and tissue imaging, with a focus on the advances and applications of CARS and SRS microscopy, for a better understanding of the metabolism and dynamics of lipids, protein, glucose, and nucleic acids in mammalian cells and tissues.

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Multiplex coherent anti-Stokes Raman scattering highlights state of chromatin condensation in CH region

Cited by 28 publications

References 42 publications

Multiplex coherent anti-Stokes Raman scattering microspectroscopy detection of lipid droplets in cancer cells expressing TrkB

Multiplex coherent anti-Stokes Raman scattering microspectroscopy detection of lipid droplets in cancer cells expressing TrkB

Recent Advances in the Synthesis of Intra‐Nanogap Au Plasmonic Nanostructures for Bioanalytical Applications

Mammalian cell and tissue imaging using Raman and coherent Raman microscopy

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