Resonance Raman Probes for Organelle-Specific Labeling in Live Cells

Kuzmin, A.; Pliss, Artem; Lim, Chang Keun; Heo, Jeongyun; Kim, Sehoon; Rzhevskii, Alexander; Gu, Bobo; Yong, Ken‐Tye; Wen, Shangchun; Prasad, Paras N.

doi:10.1038/srep28483

Cited by 37 publications

(43 citation statements)

References 33 publications

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“…These scaffolds have extremely low fluorescence and were modified to have red-shifted absorption for resonant Raman measurement. 66 And cell membrane, mitochondria and lysosome anchoring motifs were covalently linked to the quencher to allow organelle-specific imaging by resonant Raman imaging in live cells, although with limited imaging speed and sensitivity.…”

Section: Organelle Imagingmentioning

confidence: 99%

Applications of vibrational tags in biological imaging by Raman microscopy

Zhao

Shen

et al. 2017

Analyst

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As a superb tool to visualize and study the spatial-temporal distribution of chemicals, Raman microscopy has made big impacts to many disciplines of science. While the label-free imaging has been the prevailing strategy in Raman microscopy, recent development and applications of vibrational/Raman tags, particularly when coupled with stimulated Raman Scattering (SRS) microscopy, have generated intense excitement in biomedical imaging. SRS imaging of vibrational tags has enabled researchers to study a wide range of small biomolecules with high specificity, sensitivity and multiplex capability, at single live cell level, tissue level or even in vivo. As reviewed in the article, this platform has facilitated imaging distribution and dynamics of small molecules such as glucose, lipids, amino acids, nucleic acids, and drugs that are otherwise difficult to do with other means. As both the vibrational tags and Raman instrumental development progress rapidly and synergistically, we anticipate that the technique will shed light onto an even broader spectrum of biomedical problems.

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Section: Organelle Imagingmentioning

confidence: 99%

Applications of vibrational tags in biological imaging by Raman microscopy

Zhao

Shen

et al. 2017

Analyst

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“…In particular, Raman microspectroscopy has benefited from technological advances that have enabled biological samples to be analysed with improved signal or more suitable measurement speed 35 . These include Resonance Raman Scattering (RRS) 36 , Surface-Enhanced Raman Scattering (SERS) 37 , and Coherent Anti-Stokes Raman Scattering (CARS) 38 , 39 . A comprehensive overview of the advances in nonlinear Raman microspectroscopy con be found in the recent literature 40 .…”

Section: Introductionmentioning

confidence: 99%

Classification of M1/M2-polarized human macrophages by label-free hyperspectral reflectance confocal microscopy and multivariate analysis

Bertani

Mozetic

Fioramonti

et al. 2017

Sci Rep

165

126

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The possibility of detecting and classifying living cells in a label-free and non-invasive manner holds significant theranostic potential. In this work, Hyperspectral Imaging (HSI) has been successfully applied to the analysis of macrophagic polarization, given its central role in several pathological settings, including the regulation of tumour microenvironment. Human monocyte derived macrophages have been investigated using hyperspectral reflectance confocal microscopy, and hyperspectral datasets have been analysed in terms of M1 vs. M2 polarization by Principal Components Analysis (PCA). Following PCA, Linear Discriminant Analysis has been implemented for semi-automatic classification of macrophagic polarization from HSI data. Our results confirm the possibility to perform single-cell-level in vitro classification of M1 vs. M2 macrophages in a non-invasive and label-free manner with a high accuracy (above 98% for cells deriving from the same donor), supporting the idea of applying the technique to the study of complex interacting cellular systems, such in the case of tumour-immunity in vitro models.

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“…21 Along a similar spectroscopic line but with probe engineering, recent utilization of a special class of probes, fluorescent quenchers with extremely low quantum yield, as resonance Raman reporters demonstrated specific organelle imaging capability by spontaneous Raman microscopy. 22,23 Compared to nonresonant Raman (i.e., ω exc ≪ ω 0 ), it showed 3 orders of magnitude signal enhancement. 22,23 Such a Raman signal boost and fluorescent background reduction from resonance Raman reporters made spontaneous Raman imaging of certain targets possible in living cells.…”

mentioning

confidence: 99%

“…22,23 Compared to nonresonant Raman (i.e., ω exc ≪ ω 0 ), it showed 3 orders of magnitude signal enhancement. 22,23 Such a Raman signal boost and fluorescent background reduction from resonance Raman reporters made spontaneous Raman imaging of certain targets possible in living cells. However, its sensitivity and imaging speed are still unsatisfying for visualizing a wide variety of biomolecules.…”

mentioning

confidence: 99%

Electronic Preresonance Stimulated Raman Scattering Microscopy

Min

2018

J. Phys. Chem. Lett.

116

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Optical microscopy has generated great impact for modern research. While fluorescence microscopy provides the ultimate sensitivity, it generally lacks chemical information. Complementarily, vibrational imaging methods provide rich chemical-bond-specific contrasts. Nonetheless, they usually suffer from unsatisfying sensitivity or compromised biocompatibility. Recently, electronic preresonance stimulated Raman scattering (EPR-SRS) microscopy was reported, achieving simultaneous high detection sensitivity and superb vibrational specificity of chromophores. With newly synthesized Raman-active dyes, this method readily breaks the optical color barrier of fluorescence microscopy and is well-suited for supermultiplex imaging in biological samples. In this Perspective, we first review previous utilizations of electronic resonance in various Raman spectroscopy and microscopy. We then discuss the physical origin and uniqueness of the electronic preresonance region, followed by quantitative analysis of the enhancement factors involved in EPR-SRS microscopy. On this basis, we provide an outlook for future development as well as the broad applications in biophotonics.

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Resonance Raman Probes for Organelle-Specific Labeling in Live Cells

Cited by 37 publications

References 33 publications

Applications of vibrational tags in biological imaging by Raman microscopy

Applications of vibrational tags in biological imaging by Raman microscopy

Classification of M1/M2-polarized human macrophages by label-free hyperspectral reflectance confocal microscopy and multivariate analysis

Electronic Preresonance Stimulated Raman Scattering Microscopy

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