Super resolution measurement of collagen fibers in biological samples: Validation of a commercial solution for multiphoton microscopy

Barlow, Aaron; Mostaço-Guidolin, Leila B.; Osei, Emmanuel T.; Booth, Steven; Hackett, Tillie-Louise

doi:10.1371/journal.pone.0229278

Cited by 19 publications

(12 citation statements)

References 40 publications

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“…Imaging and cellular analysis of airway fibroblasts. Collagen I gels were fixed in 4% paraformaldehyde, stained with 4' ,6-diamidino-2-phenylindole (DAPI) to identify nuclei, Phalloidin (Thermo-Fisher Scientific, Waltham, USA) for F-actin, and E-cadherin using a rabbit monoclonal (DECMA-1) antibody (Abcam, ab11512), then imaged at 20x magnification using a Zeiss LSM-880 inverted confocal microscope (Carl-Zeiss, Germany), with a Coherent Chameleon Ultra II femtosecond Ti:sapphire laser (Coherent, CA) 54 . The cell area and cell numbers of PAFs within collagen gels were determined with Fiji as previously described 23 .…”

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confidence: 99%

Epithelial-interleukin-1 inhibits collagen formation by airway fibroblasts: Implications for asthma

Osei

Mostaço-Guidolin

Hsieh

et al. 2020

Sci Rep

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In asthma, the airway epithelium has an impaired capacity to differentiate and plays a key role in the development of airway inflammation and remodeling through mediator release. The study objective was to investigate the release of (IL)-1 family members from primary airway epithelial-cells during differentiation, and how they affect primary airway fibroblast (PAF)-induced inflammation, extracellular matrix (ECM) production, and collagen I remodeling. The release of IL-1α/β and IL-33 during airway epithelial differentiation was assessed over 20-days using air-liquid interface cultures. The effect of IL-1 family cytokines on airway fibroblasts grown on collagen-coated well-plates and 3-dimensional collagen gels was assessed by measurement of inflammatory mediators and ECM proteins by ELISA and western blot, as well as collagen fiber formation using non-linear optical microscopy after 24-hours. The production of IL-1α is elevated in undifferentiated asthmatic-PAECs compared to controls. IL-1α/β induced fibroblast pro-inflammatory responses (CXCL8/IL-8, IL-6, TSLP, GM-CSF) and suppressed ECMproduction (collagen, fibronectin, periostin) and the cell's ability to repair and remodel fibrillar collagen I via LOX, LOXL1 and LOXL2 activity, as confirmed by inhibition with β-aminopropionitrile. These data support a role for epithelial-derived-IL-1 in the dysregulated repair of the asthmatic-EMTU and provides new insights into the contribution of airway fibroblasts in inflammation and airway remodeling in asthma.

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Section: See Online Supplement For Full Methods Sample Collection Pmentioning

confidence: 99%

Epithelial-interleukin-1 inhibits collagen formation by airway fibroblasts: Implications for asthma

Osei

Mostaço-Guidolin

Hsieh

et al. 2020

Sci Rep

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“…In summary, we have described a thorough theoretical description of SHG‐PR. Existing, standard SHG microscope can readily be transformed into SHG‐PR systems by replacing the point detector with a position‐sensitive camera or Airyscan detector for SHG signal detection (Barlow et al ., 2020). Then, a whole image for the excited region will be recorded for each position of the scanning focus.…”

Section: Discussionmentioning

confidence: 99%

Second harmonic generation microscopy using pixel reassignment

Wang

Zhang

et al. 2020

Journal of Microscopy

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Second harmonic generation (SHG) microscopy is expected to be a powerful tool for observing the cellular-level functionality and morphology information of thick tissue owe to its unique imaging properties. However, the maximum attainable resolution obtainable by SHG microscopy is limited by the use of long-wavelength, near-infrared excitation. In this paper, we report the use of pixel reassignment to improve the spatial resolution of SHG microscopy. The SHG signal is imaged onto a position-sensitive camera, instead of a point detector typically used in conventional SHG microscope. The data processing is performed through pixel reassignment and subsequent deblurring operation. We present the basic principle and a rigorous theoretical model for SHG microscopy using pixel reassignment (SHG-PR). And for the first time, the optimal reassignment factor for SHG-PR is derived based on the coherent characteristics and the dependence of wavelength in SHG microscopy. To evaluate the spatial resolution improvement, images of nano-beads separated by different distances and of a microtubule array have been simulated. We gain about a 1.5-fold spatial resolution enhancement compared to conventional SHG microscopy. When a further deblurring operation is implemented, this method allows for a total spatial resolution enhancement of about 1.87. Additionally, we demonstrate the validity of SHG-PR for raw data with noise.

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“…A recent study by Barlow et al has shown use of the Zeiss Airyscan (a 32-channel GaAsP photomultiplier tube (PMT) array detector (17)) on top of MPM and SHG imaging can improve resolution up by a factor of 1.7× (18).…”

Section: Introductionmentioning

confidence: 99%

tAF-MUSICAL: Autofluorescence Super-resolution Microscopy for Molecular Histopathology of Matrix Proteins in Fibrotic Diseases

Ghosh

Chatterjee²,

Paul

et al. 2020

Preprint

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Fibrosis is an extracellular matrix disease affecting several vital organs' functions and can lead to life-threatening pathologies like cancer. The standard diagnostic protocol involves an immuno-histochemical examination of the fibrosis-linked protein's distribution in the tissues. Of specific interest are the primarily affected dense matrix-proteins like collagen. But labeling such dense proteins is prone to subjectivity. Besides distribution, the collagen nano-fibril characteristics, usually revealed by ultra-structural imaging, also have diagnostic relevance. Super-resolution microscopy can aid high-resolution clinical decisions by imaging protein nano-structures but is challenging for dense proteins. Here we exploit the natural fluctuations of tissue autofluorescence (tAF) signal from proteins in dense tissue matrix for super-resolving them. We achieved super-resolution over a clinically relevant large area using a simple, low-cost, low numerical aperture (NA) optical microscope and a recent fluctuation-based nanoscopy technique MUSICAL. We could quantify protein distribution and nano-fibril dimensions as low as 43 nm in human oral mucosa with pre-cancer fibrosis and mouse skin samples to assess disease progression. Thus tAF-MUSICAL enables early, label-free, and high-resolution diagnosis of matrix-associated diseases like fibrosis.

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Super resolution measurement of collagen fibers in biological samples: Validation of a commercial solution for multiphoton microscopy

Cited by 19 publications

References 40 publications

Epithelial-interleukin-1 inhibits collagen formation by airway fibroblasts: Implications for asthma

Epithelial-interleukin-1 inhibits collagen formation by airway fibroblasts: Implications for asthma

Second harmonic generation microscopy using pixel reassignment

tAF-MUSICAL: Autofluorescence Super-resolution Microscopy for Molecular Histopathology of Matrix Proteins in Fibrotic Diseases

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