Organelle specific imaging in live cells and immuno-labeling using resonance Raman probe

Li, Yuee; Heo, Jeongyun; Lim, Chang Keun; Pliss, Artem; Kachynski, Aliaksandr V.; Kuzmin, A.; Kim, Sehoon; Prasad, Paras N.

doi:10.1016/j.biomaterials.2015.02.056

Cited by 42 publications

(46 citation statements)

References 22 publications

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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.…”

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“…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.…”

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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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Electronic Preresonance Stimulated Raman Scattering Microscopy

Min

2018

J. Phys. Chem. Lett.

116

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“…A major obstacle of this technique for subcellular analysis is that the location of specific organelles should be resolved to target acquisition of Raman spectra and analyze the content of a single organelle. Meanwhile, subcellular compartments, with a partial exception of mitochondria114, do not exhibit organelle-specific vibrational bands and, therefore, cannot be recognized by the Raman imaging of intrinsic cellular components.…”

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Resonance Raman Probes for Organelle-Specific Labeling in Live Cells

Kuzmin

Pliss

Lim

et al. 2016

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Raman microspectroscopy provides for high-resolution non-invasive molecular analysis of biological samples and has a breakthrough potential for dissection of cellular molecular composition at a single organelle level. However, the potential of Raman microspectroscopy can be fully realized only when novel types of molecular probes distinguishable in the Raman spectroscopy modality are developed for labeling of specific cellular domains to guide spectrochemical spatial imaging. Here we report on the design of a next generation Raman probe, based on BlackBerry Quencher 650 compound, which provides unprecedentedly high signal intensity through the Resonance Raman (RR) enhancement mechanism. Remarkably, RR enhancement occurs with low-toxic red light, which is close to maximum transparency in the biological optical window. The utility of proposed RR probes was validated for targeting lysosomes in live cultured cells, which enabled identification and subsequent monitoring of dynamic changes in this organelle by Raman imaging.

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“…For this reason, UV-absorbing small aromatic molecules without fluorescence emission have usually been studied as RR markers [8,9]. Recently, we have reported a new generation of non-UV RR marker based on a non-fluorescent small molecule (azobenzene) that produces Raman enhancement under non-toxic visible light excitation (instead of phototoxic UV irradiation) to enable target-specific imaging in live cells and immunolabeling [10]. In that report, cellular target specificity was endowed by chemical conjugation of the RR-active molecule with organelle-binding ligands or antibodies.…”

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confidence: 99%