Stimulated Raman scattering of polymer nanoparticles for multiplexed live-cell imaging

Hu, Fanghao; Brucks, Spencer D.; Lambert, Tristan H.; Campos, Luis M.; Min, Wei

doi:10.1039/c7cc01860f

Cited by 42 publications

(38 citation statements)

References 37 publications

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“…SRS imaging is an emerging technique for evaluating nanoparticle dynamics at the cellular level. Recently, Hu et al demonstrated that SRS imaging could be used in tandem with bio-orthogonal labelling of styrene-based nanoparticles, to enable multiplex detection of three Raman-active nanoparticle species in living HeLa cells [90]. Raman-active monomers incorporating alkyne, nitrile, and C–D groups were used to prepare Raman-active polymer dots (Figure 7).…”

Section: Coherent Raman Imagingmentioning

confidence: 99%

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Raman Imaging of Nanocarriers for Drug Delivery

et al. 2019

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The efficacy of pharmaceutical agents can be greatly improved through nanocarrier delivery. Encapsulation of pharmaceutical agents into a nanocarrier can enhance their bioavailability and biocompatibility, whilst also facilitating targeted drug delivery to specific locations within the body. However, detailed understanding of the in vivo activity of the nanocarrier-drug conjugate is required prior to regulatory approval as a safe and effective treatment strategy. A comprehensive understanding of how nanocarriers travel to, and interact with, the intended target is required in order to optimize the dosing strategy, reduce potential off-target effects, and unwanted toxic effects. Raman spectroscopy has received much interest as a mechanism for label-free, non-invasive imaging of nanocarrier modes of action in vivo. Advanced Raman imaging techniques, including coherent anti-Stokes Raman scattering (CARS) and stimulated Raman scattering (SRS), are paving the way for rigorous evaluation of nanocarrier activity at the single-cell level. This review focuses on the development of Raman imaging techniques to study organic nanocarrier delivery in cells and tissues.

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Section: Coherent Raman Imagingmentioning

confidence: 99%

“…The polymer dots are detected at discrete frequencies in the HeLa cells: ( c ) C≡C dot (2163 cm −1 ), ( d ) C≡N dot (2232 cm −1 ), and ( e ) C–D dot (2293 cm −1 ), with the cellular contrast detected at 2845 cm −1 (CH 2 , lipids) in the merge images. Reproduced with permission from [90], Copyright Royal Society of Chemistry, 2017.…”

Section: Figurementioning

confidence: 99%

Raman Imaging of Nanocarriers for Drug Delivery

et al. 2019

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“…[14][15][16] As label-free imaging methods, both CARS and SRS have been successfully used in a broad spectrum of applications, including the molecular imaging of cell metabolites, [17,18] real-time monitoring of drug delivery, [19] differentiating tumor margins from healthy tissues, [20] and visualizing specially designed polymer dots in cells via various Raman tags. [21] With the latest technical advances in hyperspectral SRS, the chemical mapping of molecules and thus the metabolic fingerprinting of multiple molecular species have become possible. [19,[22][23][24] Nevertheless, direct visualization and quantification of dissimilar NPs in single cells by hyperspectral SRS have not been evaluated.…”

Section: Doi: 101002/smll201703246mentioning

confidence: 99%

“…Coherent anti‐Stokes RS (CARS) and stimulated RS (SRS) overcome these problems by stimulating the Raman transition of biomolecules via nonlinear interactions with two coherent pulse laser beams . As label‐free imaging methods, both CARS and SRS have been successfully used in a broad spectrum of applications, including the molecular imaging of cell metabolites, real‐time monitoring of drug delivery, differentiating tumor margins from healthy tissues, and visualizing specially designed polymer dots in cells via various Raman tags . With the latest technical advances in hyperspectral SRS, the chemical mapping of molecules and thus the metabolic fingerprinting of multiple molecular species have become possible .…”

Section: Introductionmentioning

confidence: 99%

Label‐Free Imaging of Nanoparticle Uptake Competition in Single Cells by Hyperspectral Stimulated Raman Scattering

Bin

Yan

Xiao

et al. 2017

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Imaging and quantification of nanoparticles in single cells in their most natural condition are expected to facilitate the biotechnological applications of nanoparticles and allow for better assessment of their biosafety risks. However, current imaging modalities either require tedious sample preparation or only apply to nanoparticles with specific physicochemical characteristics. Here, the emerging hyperspectral stimulated Raman scattering (SRS) microscopy, as a label-free and nondestructive imaging method, is used for the first time to investigate the subcellular distribution of nanoparticles in the protozoan Tetrahymena thermophila. The two frequently studied nanoparticles, polyacrylate-coated α-Fe O and TiO , are found to have different subcellular distribution pattern as a result of their dissimilar uptake routes. Significant uptake competition between these two types of nanoparticles is further discovered, which should be paid attention to in future bioapplications of nanoparticles. Overall, this study illustrates the great promise of hyperspectral SRS as an analytical imaging tool in nanobiotechnology and nanotoxicology.

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“…Over the past years, the number of techniques available to study cell uptake and intracellular trafficking of polymers and polymer nanoparticles has increased and now also includes several methods that do not require the use of fluorescently labeled polymers. 15 Raman spectroscopy, for example, uses vibrational labels, often isotopes, 16−19 and nanoscale secondary ion mass spectrometry (NanoSIMS) allows the intracellular tracking of trace elements or isotopes. 20−22 Synchrotron X-ray fluorescence imaging has also been used to visualize elements and metals in cellular environments.…”

Section: Introductionmentioning

confidence: 99%

Synthesis and Postpolymerization Modification of Fluorine-End-Labeled Poly(Pentafluorophenyl Methacrylate) Obtained via RAFT Polymerization

et al. 2018

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Chain-end-labeled polymers are interesting for a range of applications. In polymer nanomedicine, chain-end-labeled polymers are useful to study and help understand cellular internalization and intracellular trafficking processes. The recent advent of fluorescent label-free techniques, such as nanoscale secondary ion mass spectrometry (NanoSIMS), provides access to high-resolution intracellular mapping that can complement information obtained using fluorescent-labeled materials and confocal microscopy and flow cytometry. Using poly( N -(2-hydroxypropyl)methacrylamide) (PHPMA) as a prototypical polymer nanomedicine, this paper presents a synthetic strategy to polymers that contain trace element labels, such as fluorine, which can be used for NanoSIMS analysis. The strategy presented in this paper is based on reversible addition fragmentation chain transfer (RAFT) polymerization of pentafluorophenyl methacrylate (PFMA) mediated by two novel chain-transfer agents (CTAs), which contain either one (α) or two (α,ω) fluorine labels. In the first part of this study, via a number of polymerization experiments, the polymerization properties of the fluorinated RAFT CTAs were established. 19 F NMR spectroscopy revealed that these fluorinated RAFT agents possess unique spectral signatures, which allow to directly monitor RAFT agent conversion and measure end-group fidelity. Comparison with 4-cyanopentanoic acid dithiobenzoate, which is a standard CTA for the RAFT polymerization of PFMA, revealed that the introduction of one or two fluorine labels does not significantly affect the polymerization properties of the CTA. In the last part of this paper, a proof-of-concept study is presented that demonstrates the feasibility of the fluorine-labeled poly(pentafluorophenyl methacrylate) polymers as platforms for the postpolymerization modification to generate PHPMA-based polymer nanomedicines.

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Stimulated Raman scattering of polymer nanoparticles for multiplexed live-cell imaging

Cited by 42 publications

References 37 publications

Raman Imaging of Nanocarriers for Drug Delivery

Raman Imaging of Nanocarriers for Drug Delivery

Label‐Free Imaging of Nanoparticle Uptake Competition in Single Cells by Hyperspectral Stimulated Raman Scattering

Synthesis and Postpolymerization Modification of Fluorine-End-Labeled Poly(Pentafluorophenyl Methacrylate) Obtained via RAFT Polymerization

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