Quantifying single-cell secretion in real time using resonant hyperspectral imaging

Juan-Colás, José; Hitchcock, Ian S.; Coles, Mark; Johnson, Steven; Krauss, Thomas F.

doi:10.1073/pnas.1814977115

Cited by 35 publications

(32 citation statements)

References 18 publications

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“…Flow cytometry [114,116] and droplet microfluidics [131] are already able to pick up heterogeneity, but only at the microscopic scale. We therefore see exciting opportunities for techniques such as SPRi [51] or GMR imaging [32,79] to add their nanoscale sensing ability to this problem.…”

Section: Discussionmentioning

confidence: 99%

“…These results clearly demonstrate that the metasurface approach can be used for nanoscopic analysis because the refractive index being probed is that of the bacterial membrane. This ability could open up a variety of studies including, for example, cell adhesion and secretion monitoring that have previously only been demonstrated with mammalian cells [79,80], but have never been applied to bacteria. Monitoring the response of bacteria to antibiotic challenge is another promising area of research opened up by these structures, which is discussed in more detail in Section 3.…”

Section: Resonant Metasurfacesmentioning

confidence: 99%

“…Interestingly, some of the techniques are only nanoscopic in one dimension, that is they probe the bacterium within a few hundred nanometres of the surface but average over an illuminated area of 100s of μm or even millimetres, whereas others are much more localized and are able to probe individual bacteria. SPRi or hyperspectral imaging with GMRs [20,49,79], fall into the latter category.…”

Section: Evanescent Wave-based Astsmentioning

confidence: 99%

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Nanophotonics for bacterial detection and antimicrobial susceptibility testing

2020

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Photonic biosensors are a major topic of research that continues to make exciting advances. Technology has now improved sufficiently for photonics to enter the realm of microbiology and to allow for the detection of individual bacteria. Here, we discuss the different nanophotonic modalities used in this context and highlight the opportunities they offer for studying bacteria. We critically review examples from the recent literature, starting with an overview of photonic devices for the detection of bacteria, followed by a specific analysis of photonic antimicrobial susceptibility tests. We show that the intrinsic advantage of matching the optical probed volume to that of a single, or a few, bacterial cell, affords improved sensitivity while providing additional insight into single-cell properties. We illustrate our argument by comparing traditional culture-based methods, which we term macroscopic, to microscopic free-space optics and nanoscopic guided-wave optics techniques. Particular attention is devoted to this last class by discussing structures such as photonic crystal cavities, plasmonic nanostructures and interferometric configurations. These structures and associated measurement modalities are assessed in terms of limit of detection, response time and ease of implementation. Existing challenges and issues yet to be addressed will be examined and critically discussed.

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Section: Discussionmentioning

confidence: 99%

Section: Resonant Metasurfacesmentioning

confidence: 99%

Section: Evanescent Wave-based Astsmentioning

confidence: 99%

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Nanophotonics for bacterial detection and antimicrobial susceptibility testing

2020

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“…Two major aspects have so far been missed, however. a) The dynamic range of the resonance is essential for a high signal-to-noise measurement, yet much of the previous work has aimed to maximise the Q-factor alone; b) The high sensitivity can also be most fruitfully applied to imaging applications, which so far has been limited to mammalian cells due to their larger (5-10 µm) size [9,20,21]. A study aiming to concurrently optimize both the imaging and the sensing function is still missing.…”

Section: Introductionmentioning

confidence: 99%

Dielectric Nanohole Array Metasurface For High-Resolution Near-Field Sensing and Imaging

Conteduca

Barth

Pitruzzello

et al. 2020

Preprint

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Dielectric metasurfaces support resonances that are widely explored both for far-field wavefront shaping and for near-field sensing and imaging. Their design explores the interplay between localised Mie resonances and extended Bragg resonances, with a typical trade-off between Q-factor and light localisation; high Q-factors are desirable for refractive index sensing while localisation is desirable for imaging resolution. Here, we show that a dielectric metasurface consisting of a nanohole array in amorphous silicon provides a favourable trade-off between these requirements. We have designed and realised the metasurface to support two optical modes both with sharp Fano resonances that exhibit relatively high Q factors and strong spatial confinement, thereby concurrently optimizing the device for both imaging and biochemical sensing. For the sensing application, we demonstrate a limit of detection (LOD) as low as 1pg/ml for Immunoglobulin G (IgG); for resonant imaging, we demonstrate a spatial resolution below 1µm and clearly resolve individual E.coli bacteria. The combined low LOD and high spatial resolution opens new opportunities for extending cellular studies into the realm of microbiology, e.g. for studying antimicrobial susceptibility.

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“…Recent studies approach this challenge in two ways, focusing either on maximizing detection sensitivity, or on tailoring the ideal functional-tissue models. The first approach targets real-time analysis and quantification of secreted cytokines using ultrasensitive detection method or imaged-based high-resolution spatiotemporal techniques ( Shirasaki et al, 2015 ; Juan-Colás et al, 2018 ; Saxena et al, 2018 ). Here however, because the techniques involved rely on single-cell secretion, this approach cannot probe cytokine secretion by functional tissues.…”

Section: Introductionmentioning

confidence: 99%

Microphysiological sensing platform for an in-situ detection of tissue-secreted cytokines

Hernández-Albors

Castaño

Fernández-Garibay

et al. 2019

Biosensors and Bioelectronics: X

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Understanding the protein-secretion dynamics from single, specific tissues is critical toward the advancement of disease detection and treatments. However, such secretion dynamics remain difficult to measure in vivo due to the uncontrolled contributions from other tissue populations. Here, we describe an integrated platform designed for the reliable, near real-time measurements of cytokines secreted from an in vitro single-tissue model. In our setup, we grow 3D biomimetic tissues to discretize cytokine source, and we separate them from a magnetic microbead-based biosensing system using a Transwell insert. This design integrates physiochemically controlled biological activity, high-sensitivity protein detection (LOD < 20 pg mL −1 ), and rapid protein diffusion to enable non-invasive, near real-time measurements. To showcase the specificity and sensitivity of the system, we use our setup to probe the inflammatory process related to the protein Interleukine 6 (IL-6) and to the Tumor Necrosis Factor (TNF-α). We show that our setup can monitor the time-dependence profile of IL-6 and TNF-α secretion that results from the electrical and chemical stimulation of 3D skeletal muscle tissues. We demonstrate a novel and affordable methodology for discretizing the secretion kinetics of specific tissues for advancing metabolic-disorder studies and drug-screening applications.

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Quantifying single-cell secretion in real time using resonant hyperspectral imaging

Cited by 35 publications

References 18 publications

Nanophotonics for bacterial detection and antimicrobial susceptibility testing

Nanophotonics for bacterial detection and antimicrobial susceptibility testing

Dielectric Nanohole Array Metasurface For High-Resolution Near-Field Sensing and Imaging

Microphysiological sensing platform for an in-situ detection of tissue-secreted cytokines

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