Methyl methacrylate embedding to study the morphology and immunohistochemistry of adult guinea pig and mouse cochleae

Bakó, Péter; Bassiouni, Mohamed; Eckhard, Andreas H.; Gerlinger, Imre; Frick, Claudia; Löwenheim, Hubert; Müller, Marcus

doi:10.1016/j.jneumeth.2015.07.017

Cited by 16 publications

(12 citation statements)

References 25 publications

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“…The animals were euthanized 21 days after AG application, while under deep anesthesia using an overdose of Narcoren (4 mL/kg). Then, the cochlear samples were collected, preserved, permeabilized, and blocked, as described previously 84 . The cochlear tissues were stained to detect the CtBP2 (C-terminal-binding protein 2), as described in Supplementary Information.…”

Section: Methodsmentioning

confidence: 99%

Lower ototoxicity and absence of hidden hearing loss point to gentamicin C1a and apramycin as promising antibiotics for clinical use

Ishikawa

García-Mateo

Čusak

et al. 2019

Sci Rep

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Spread of antimicrobial resistance and shortage of novel antibiotics have led to an urgent need for new antibacterials. Although aminoglycoside antibiotics (AGs) are very potent anti-infectives, their use is largely restricted due to serious side-effects, mainly nephrotoxicity and ototoxicity. We evaluated the ototoxicity of various AGs selected from a larger set of AGs on the basis of their strong antibacterial activities against multidrug-resistant clinical isolates of the ESKAPE panel: gentamicin, gentamicin C1a, apramycin, paromomycin and neomycin. Following local round window application, dose-dependent effects of AGs on outer hair cell survival and compound action potentials showed gentamicin C1a and apramycin as the least toxic. Strikingly, although no changes were observed in compound action potential thresholds and outer hair cell survival following treatment with low concentrations of neomycin, gentamicin and paromomycin, the number of inner hair cell synaptic ribbons and the compound action potential amplitudes were reduced. This indication of hidden hearing loss was not observed with gentamicin C1a or apramycin at such concentrations. These findings identify the inner hair cells as the most vulnerable element to AG treatment, indicating that gentamicin C1a and apramycin are promising bases for the development of clinically useful antibiotics.

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Section: Methodsmentioning

confidence: 99%

Lower ototoxicity and absence of hidden hearing loss point to gentamicin C1a and apramycin as promising antibiotics for clinical use

Ishikawa

García-Mateo

Čusak

et al. 2019

Sci Rep

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“…While the use of DAPI‐oil is fast and convenient, we observed that even with anti‐fading solution (Vectashield) bleaching of the dye occurred relatively quickly, by observation with UV‐light (∼ 5 min). An aqueous staining approach followed by embedding in anti‐fading solution as reported for methacrylate embedded tissue might provide better results in cases where longer observation times are required (Bako et al ).…”

Section: Assessmentmentioning

confidence: 99%

Hard and soft plastic resin embedding for single‐cell element uptake investigations of marine‐snow‐associated microorganisms using nano‐scale secondary ion mass spectrometry

Rogge

Flintrop

Iversen

et al. 2018

Limnology & Ocean Methods

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Marine snow aggregates are microhabitats for diverse microbial communities with various active metabolic pathways. Rapid recycling and symbiotic transfer of nutrients within aggregates poses a significant challenge for accurately assessing aggregate-associated turnover rates. Although single-cell uptake measurements are well-established for free-living microorganisms, suitable methods for cells embedded in marine snow are currently lacking. Comparable cell-specific measurements within sinking pelagic aggregates would have the potential to address core questions regarding aggregate-associated fluxes. However, the capacity to perform microscale studies is limited by the difficulty of sampling and preserving the fragile aggregate structure. Furthermore, the application of nano-scale secondary ion mass spectrometry (NanoSIMS) to aggregates is complicated by technical requirements related to vacuum and ablation resistance. Here, we present a NanoSIMSoptimized method for fixation, embedding, and sectioning of marine snow. Stable isotope labeling of laboratory-generated aggregates enabled visualization of label incorporation into prokaryotic and eukaryotic cells embedded in the aggregate structure. The current method is also amenable to various staining procedures, including transparent exopolymer particles, Coomassie stainable particles, nucleic acids, and eukaryotic cytoplasm. We demonstrate the potential for using structural stains to generate three-dimensional (3D) models of marine snow and present a simplified calculation of porosity and fractal dimension. This multipurpose method enables combined investigations of 3D aggregate structure, spatial microbial distribution, and single-cell activity within individual aggregates and provides new possibilities for future studies on microbial interactions and elemental uptake within marine snow.

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“…A modification of the previously shown embedding method using a novel polymerization system was used. 29,30 The polymerization system is based on poly(methyl methacrylate). Polymerization occurs in the absence of oxygen and at low temperatures (−2 • C to −20 • C) and enables to perform immunohistochemical staining for hard tissues.…”

Section: Embedding Of Tissue Culture Samplesmentioning

confidence: 99%

Zirconia implants with improved attachment to the gingival tissue

Shahramian

Gasik

Kangasniemi

et al. 2020

Journal of Periodontology

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Background Gingival tissue attachment is known to be important for long‐term prognosis of implants. This in vitro study evaluated the gingival attachment to zirconia implants and zirconia implants modified with sol‐gel derived TiO2 coatings. Methods Zirconia endodontic posts (n = 23) were used to function as implants that were inserted into the center of full‐thickness porcine gingival explants (n = 31). The tissue/implant specimens were then individually placed at an air/liquid interface on a stainless‐steel grid in cell culture wells containing a nutrient solution. The tissue cultures were incubated at 37°C in a 5% CO2 environment and at days 7 and 14, the specimens were harvested and analyzed by dynamic mechanical analysis (DMA) measurements under dynamic loading conditions mimicking natural mastication. Specimens were also analyzed by immunohistochemical staining identifying the laminin (Ln) γ2 chain specific for Ln‐332, which is known to be a crucial molecule for the proper attachment of epithelium to tooth/implant surface. Results Tissue attachment to TiO2‐coated zirconia demonstrated higher dynamic modulus of elasticity and higher creep modulus, meaning that the attachment is stronger and more resistant to damage during function over time. Laminin γ2 was identified in the attachment of epithelium to TiO2‐coated zirconia. Conclusions Both DMA and histological analysis support each other, so the gingival tissue is more strongly attached to sol‐gel derived TiO2‐coated zirconia than uncoated zirconia. Immunohistochemical staining showed that TiO2 coating may enhance the synthesis and deposition of Ln‐332 in the epithelial attachment to the implant surface.

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Methyl methacrylate embedding to study the morphology and immunohistochemistry of adult guinea pig and mouse cochleae

Abstract: Our results showed that the Technovit 9100 New(®) embedding system provided highly preserved morphology and immunogenicity with our protocol in adult guinea pig and mouse cochleae.

Cited by 16 publications

References 25 publications

Lower ototoxicity and absence of hidden hearing loss point to gentamicin C1a and apramycin as promising antibiotics for clinical use

Lower ototoxicity and absence of hidden hearing loss point to gentamicin C1a and apramycin as promising antibiotics for clinical use

Hard and soft plastic resin embedding for single‐cell element uptake investigations of marine‐snow‐associated microorganisms using nano‐scale secondary ion mass spectrometry

Zirconia implants with improved attachment to the gingival tissue

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