Raster image correlation spectroscopy as a novel tool to study interactions of macromolecules with nanofiber scaffolds

Norris, Sam C. P.; Humpolíčková, Jana; Amler, Evžen; Huranová, Martina; Buzgo, Matěj; Macháň, Radek; Lukáš, David; Hof, Martin

doi:10.1016/j.actbio.2011.07.012

Cited by 18 publications

(21 citation statements)

References 33 publications

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“…At the bench, confocal fluorescence techniques, like fluorescence recovery after photobleaching (FRAP), have been used to measure the diffusion of fluorescent solutes in cartilage . However, FRAP has significant barriers to bedside translation, including high solute concentration (∼μM) and high laser power (∼mW) requirements, which present concerns with in vivo cytocompatibility, and potentially long diagnostic procedures due to fluorescence recovery timescales. Other photobleaching‐based assessment techniques, including continuous point photobleaching and scanning microphotolysis, have similar limitations.…”

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confidence: 99%

“…Other photobleaching‐based assessment techniques, including continuous point photobleaching and scanning microphotolysis, have similar limitations. On the other hand, fluorescence‐based correlation spectroscopy techniques are far less “intrusive,” faster, and indeed simpler, overcoming many of these limitations …”

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confidence: 99%

“…Critically, both FCS and RICS require significantly reduced solute concentrations (on the order of nanomolar) and laser powers (∼μW) for assessment of diffusivity, which makes them less invasive and cytotoxic, and more amenable to clinical application if paired with an appropriate imaging apparatus, such as a confocal arthroscope. Recent studies regarding the sensitivity of these correlation spectroscopy techniques in cartilage suggest that they may be able to detect the changes associated with disease progression.…”

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confidence: 99%

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Quantification of solute diffusivity in osteoarthritic human femoral cartilage using correlation spectroscopy

Graham

Wright

Burris³

et al. 2018

Journal Orthopaedic Research

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Osteoarthritis is a chronic joint disease characterized by articular cartilage degeneration, pain, and disability. As an avascular tissue, the movement of water and solutes through the tissue is critical to cartilage health and function, and early changes in solute diffusivity due to micro-scale changes in the properties of cartilage's extracellular matrix might precede clinical symptoms. A diagnostic technique for quantifying alteration to the diffusive environment of cartilage that precedes macroscopic changes may allow for the earlier identification of osteoarthritic disease, facilitating earlier intervention strategies. Toward this end, we used two confocal microscopy-based correlation spectroscopy techniques, fluorescence correlation spectroscopy and raster image correlation spectroscopy, to quantify the diffusion of two small solutes, fluorescein and 3k dextran, within human osteoarthritic articular cartilage. Our goal was to determine if these relatively simple optical correlation spectroscopy techniques could detect changes in solute diffusivity associated with increasing cartilage damage as assessed by International Cartilage Repair Society scoring guidelines, and if these measures are correlated with mechanical and compositional measures of cartilage health. Our data show a modest, yet significant increase in solute diffusivity and cartilage permeability with increasing osteoarthritis score (grades 0-2), with a strong correlation between diffusion coefficients, permeability, and cartilage composition. The described correlation spectroscopy techniques are quick, simple, and easily adapted to existing laboratory workflow and equipment. Furthermore, the minimal solute concentrations and laser powers required for analysis, combined with recent advances in arthroscopic microscopy, suggest correlation spectroscopy techniques as translational candidates for development into early OA diagnosis tools. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.

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Quantification of solute diffusivity in osteoarthritic human femoral cartilage using correlation spectroscopy

Graham

Wright

Burris³

et al. 2018

Journal Orthopaedic Research

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“…At the time of that work, the time resolution was not sufficient to acquire temporal FCS functions at each spot, but it was used to observe the temporal development of spatial correlations. Raster image correlation spectroscopy (RICS) uses the temporal information inherent in a scanning confocal microscope to allow the calculation of spatio-temporal correlations [24] and can be used to derive diffusion and binding kinetics [25]. However, measurements are still not simultaneous over the whole sample and require laser scanning, which illuminates the whole sample and thus is more prone to bleaching and photodamage.…”

Section: Introductionmentioning

confidence: 99%

The performance of 2D array detectors for light sheet based fluorescence correlation spectroscopy

et al. 2013

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Abstract:Single plane illumination microscopy based fluorescence correlation spectroscopy (SPIM-FCS) is a new method for imaging FCS in 3D samples, providing diffusion coefficients, transport, flow velocities and concentrations in an imaging mode. SPIM-FCS records correlation functions over a whole plane in a sample, which requires array detectors for recording the fluorescence signal. Several types of image sensors are suitable for FCS. They differ in properties such as effective area per pixel, quantum efficiency, noise level and read-out speed. Here we compare the performance of several low light array detectors based on three different technologies: (1) Single-photon avalanche diode (SPAD) arrays, (2) passive-pixel electron multiplying charge coupled device (EMCCD) and (3) active-pixel scientific-grade complementary metal oxide semiconductor cameras (sCMOS). We discuss the influence of the detector characteristics on the effective FCS observation volume, and demonstrate that light sheet based SPIM-FCS provides absolute diffusion coefficients. This is verified by parallel measurements with confocal FCS, single particle tracking (SPT), and the determination of concentration gradients in space and time. While EMCCD cameras have a temporal resolution in the millisecond range, sCMOS cameras and SPAD arrays can extend the time resolution of SPIM-FCS down to 10 μs or lower. References and links1. K. M. Berland, P. T. So, and E. Gratton, "Two-photon fluorescence correlation spectroscopy: method and application to the intracellular environment," Biophy. J. 68, 694-701 (1995). 2. P. Schwille, U. Haupts, S. Maiti, and W. W. Webb, "Molecular dynamics in living cells observed by fluorescence correlation spectroscopy with one-and two-photon excitation," Biophy. J. 77, 2251-2265 (1999). 3. R. Brock, G. Vàmosi, G. Vereb, and T. M. Jovin, "Rapid characterization of green fluorescent protein fusion proteins on the molecular and cellular level by fluorescence correlation microscopy," Proc. Natl. Acad. Sci. U.S.A. 96, 10123-10128 (1999 0-1 (2010). 53. Z. Petrásek, P. Schwille, and Z. Petrášek, "Precise measurement of diffusion coefficients using scanning fluorescence correlation spectroscopy," Biophy.

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“…The ability to distinguish these two states is extremely useful in biological systems where clustering of macromolecules can differentiate cellular processes (e.g., receptor aggregation [2][3][4] and collagen fiber organization). 5 ICS has since been extended to analyze fluorophore colocalization (cross correlation ICS), [6][7][8] dispersion and diffusion in time (spatiotemporal [9][10][11] and raster ICS), [12][13][14][15][16] and macromolecular structure. 5,17 More information about these variants can be found in several recent reviews, 6 showing the versatility of this mathematical technique.…”

Section: Introductionmentioning

confidence: 99%

Theory and practical recommendations for autocorrelation-based image correlation spectroscopy

Robertson

George

2012

J. Biomed. Opt

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Abstract. Image correlation spectroscopy (ICS) is a powerful technique for detecting arrangement of fluorophores in images. This tutorial gives background into the mathematical underpinnings of ICS, specifically image autocorrelation. The effects of various artifacts and image processing steps, including background subtraction, noise, and image morphology were examined analytically and their effects on ICS analysis modeled. A series of recommendations was built based on this analysis.

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Raster image correlation spectroscopy as a novel tool to study interactions of macromolecules with nanofiber scaffolds

Cited by 18 publications

References 33 publications

Quantification of solute diffusivity in osteoarthritic human femoral cartilage using correlation spectroscopy

Quantification of solute diffusivity in osteoarthritic human femoral cartilage using correlation spectroscopy

The performance of 2D array detectors for light sheet based fluorescence correlation spectroscopy

Theory and practical recommendations for autocorrelation-based image correlation spectroscopy

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