Hyperspectral Microscopy of Near-Infrared Fluorescence Enables 17-Chirality Carbon Nanotube Imaging

Roxbury, Daniel; Jena, Prakrit V.; Williams, R. T.; Enyedi, Balázs; Niethammer, Philipp; Marcet, S.; Verhaegen, Marc; Blais-Ouellette, Sébastien; Heller, Daniel A.

doi:10.1038/srep14167

Cited by 132 publications

(195 citation statements)

References 28 publications

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“…The concentration was determined with a UV/vis/nIR spectrophotometer (Jasco, Tokyo, Japan) using the extinction coefficient A 910 = 0.02554 L·mg −1 ·cm −1 . 35 …”

Section: Methodsmentioning

confidence: 99%

DNA–Carbon Nanotube Complexation Affinity and Photoluminescence Modulation Are Independent

Jena

Safaee

Heller

et al. 2017

ACS Appl. Mater. Interfaces

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109

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Short single-stranded DNA (ssDNA) has emerged as the natural polymer of choice for non-covalently functionalizing photoluminescent single-walled carbon nanotubes. In addition, specific empirically identified DNA sequences can be used to separate single species (chiralities) of nanotubes with exceptionally high purity. Currently, only limited general principles exist for designing DNA-nanotube hybrids amenable to separation processes, due in part to an incomplete understanding of the fundamental interactions between a DNA sequence and a specific nanotube structure, while even less is known in the design of nanotube-based sensors with determined optical properties. We therefore developed a combined experimental and analysis platform, based on time-resolved near-infrared fluorescence spectroscopy, to extract the complete set of photoluminescence parameters that characterize DNA-nanotube hybrids. Here, we systematically investigated the affinity of the d(GT)n oligonucleotide family for structurally-defined carbon nanotubes by measuring photoluminescence response of the nanotube upon oligonucleotide displacement. We found, surprisingly, that the rate of displacement of oligonucleotides is independent of the coverage on the nanotube, as inferred through intrinsic optical properties of the hybrid. The kinetics of intensity modulation are essentially single exponentials, and the time constants, which quantify the stability of DNA binding, span an order of magnitude. Surprisingly, these time constants do not depend on the intrinsic optical parameters within the hybrids, suggesting that DNA-nanotube stability is not due to increased nanotube surface coverage by DNA. Further, a principal component analysis of the excitation and emission shifts, along with intensity enhancement at equilibrium accurately identified the (8,6) nanotube as the partner chirality to (GT)6 ssDNA. Combined, the chirality-resolved equilibrium and kinetics data can guide the development of DNA-nanotube pairs with tunable stability and optical modulation. Additionally, this high-throughput optical platform could function as a primary screen for mapping the DNA-chirality recognition phase space.

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“…The concentration was determined with a UV/vis/nIR spectrophotometer (Jasco, Tokyo, Japan) using the extinction coefficient A 910 = 0.02554 L·mg −1 ·cm −1 . 35 …”

Section: Methodsmentioning

confidence: 99%

DNA–Carbon Nanotube Complexation Affinity and Photoluminescence Modulation Are Independent

Jena

Safaee

Heller

et al. 2017

ACS Appl. Mater. Interfaces

Self Cite

109

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“…After sonication, the dispersion was ultracentrifuged (Sorvall Discovery 90SE) for 30 min at 250 000 g in a fixed-angle rotor (Fiberlite F50L), and the top 80% of the supernatant was extracted. The concentration was determined with a UV/vis/nIR spectrophotometer (Jasco, Tokyo, Japan) using the extinction coefficient A 910 = 0.02554 L·mg −1 ·cm −1 40 . To remove free DNA, 100 kDa Amicon centrifuge filters (Millipore) were used.…”

Section: Methodsmentioning

confidence: 99%

Cell Membrane Proteins Modulate the Carbon Nanotube Optical Bandgap via Surface Charge Accumulation

et al. 2015

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Cell adhesion is a protein-mediated process intrinsic to most living organisms. Dysfunction in cell adhesion processes is implicated in various diseases, including thrombosis and metastatic cancers. Using an approach to resolve spectral features from cell membrane-associated photoluminescent single-walled carbon nanotubes, we found that nanotube optical bandgaps respond to the electrostatic potential of the cell surface, which corresponds to cell adhesion properties. We studied the carbon nanotube emission energy response to solution ionic potentials, which suggests sensitivity to local charge accumulation. We conclude that nanotubes respond to cell surface electrostatic potentials that are mediated by membrane proteins, which vary significantly across cell types. These findings portend the optical measurement of surface electrostatic potentials for biophysical measurements and biomedical applications.

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“…However, a new generation of NIR dyes, which allow for deep tissue penetration, minimal photo damage and low background auto fluorescence are currently being developed, including squaraine and squaraine-rotaxanes derivatives, with cross sections of 33,000 GM and molecular weights much smaller than current quantum dots, the results of early studies are proving promising for the development of novel tools for subcellular imaging [17][18][19]. Additionally, fluorescent proteins [20] and nanotube-based optical sensors [21] that are excitable in the 1000-1300 nm range are being designed to allow for imaging at greater depths than what is achievable with conventional fluorophores.…”

Section: Tracking With Novel Fluorescent Proteins/dyesmentioning

confidence: 99%

Advances in In Vivo Imaging Techniques for the Visualization and Quantification of DC-T Cell Interactions

Sathappan¹,

Panhuys²

2017

J immuno Biol

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Short CommunicationSince the first application of two photon intravital microscopy (2PM) to the study of the immune system in 1997 [1], 2PM imaging has offered several major advantages over previous techniques for study immune interactions. Such as 2D plate based culture systems, as cells exhibit significantly different migratory behavior in 2D as compared to 3D. Whilst, 3D matrigel systems allow cells to migrate in a more natural manner, they fail to adequately replicate a physiological environment which is densely packed with other cell types and contains a myriad of environmental cues such as chemokines, adhesion molecules and lymphatic flow, these factors are largely overcome by the use of ex-vivo/explant organ imaging. However, this system also has significant limitations in that cells cannot be recruited to or exit the organ of interest and due to limitations in tissue oxygenation, imaging sessions can generally only be conducted for less than one hour [2]. Therefore, the use of intravital 2PM has opened up a new era of understanding in immunology. Especially in terms of how we view the importance of the migratory behavior and the interactions that occur between cells of the immune system under both homeostatic and inflammatory conditions. Perhaps this has been most enlightening when applied to the study of the adaptive immune system and the interaction between CD4+ and CD8+ T cells with dendritic cells (DC). These studies have revealed that T cells are constantly forming non-cognate interactions with DC as they migrate through the lymph node (LN) at an average velocity of 10μm/min, allowing individual DC to briefly interact with an estimated 5000 T cells an hour [3,4]. This behavior allows for the rapid scanning of the TCR repertoire during an immune response and the swift activation of antigen specific T-cells. Initial cognate interactions consist of brief phase I interactions (~0-6 h) typically lasting 10-30 minutes each, before progression to phase II interactions where stable immune synapses are formed at ~6-18 h post-activation, a transition to phase III interactions then occurs between ~18-30 h, which is typified by multiple short lived interactions and a pattern of swarming behavior [5,6]. The quality or quantity of the cognate antigen and both the phenotype and activation status of the stimulatory DC have all been shown to influence the process of activation and differentiation [7,8], and the application of novel techniques for probing this process will undoubtedly allow for a much greater understanding of the adaptive immune response. Tracking with Novel Fluorescent Proteins/Dyes:The capabilities and application of intravital 2PM for the analysis of cellular processes and the dynamics of interactions has seen an exponential growth with the introduction of several novel fluorescent proteins, optical markers and NIR (Near Infrared) fluorophores. The use of optical markers such as photoactivatable (PA) [9] fluorescent proteins PA-GFP and photo-convertible fluorophores (e.g. Kaede [10] and kikGR [1...

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Hyperspectral Microscopy of Near-Infrared Fluorescence Enables 17-Chirality Carbon Nanotube Imaging

Cited by 132 publications

References 28 publications

DNA–Carbon Nanotube Complexation Affinity and Photoluminescence Modulation Are Independent

DNA–Carbon Nanotube Complexation Affinity and Photoluminescence Modulation Are Independent

Cell Membrane Proteins Modulate the Carbon Nanotube Optical Bandgap via Surface Charge Accumulation

Advances in In Vivo Imaging Techniques for the Visualization and Quantification of DC-T Cell Interactions

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