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Related Concept Videos

Biofilms01:29

Biofilms

Biofilms are complex communities of microorganisms encased in a self-produced extracellular polysaccharide matrix attached to surfaces. These microbial consortia can include single or multiple species, providing enhanced survival benefits by forming organized, multilayered structures.The formation of biofilms occurs through four key stages: attachment, colonization, development, and dispersal.During attachment, free-swimming planktonic cells adhere to a surface, often facilitated by...

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An Analytical Tool-box for Comprehensive Biochemical, Structural and Transcriptome Evaluation of Oral Biofilms Mediated by Mutans Streptococci
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Quantitative and Structural Characterization of Streptococcus mutans Biofilms on Dental Restorative Materials Using

Krishna Kumar Kungumaraj1, Sivashankari Rajasekaran1, Bao Huynh1

  • 1Department of Biomaterial and Biomedical Sciences, Oregon Health & Science University - School of Dentistry, Portland, Oregon, USA.

Journal of Biomedical Materials Research. Part B, Applied Biomaterials
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PubMed
Summary

Standardized biofilm fixation protocols using glutaraldehyde and ethanol dehydration improve scanning electron microscopy (SEM) imaging quality for dental materials. This enhances bacterial cell and biofilm matrix discrimination in cross-section analysis.

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Concurrent Quantification of Cellular and Extracellular Components of Biofilms
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Concurrent Quantification of Cellular and Extracellular Components of Biofilms

Published on: December 10, 2013

Area of Science:

  • Dental Materials Science
  • Microbiology
  • Microscopy Techniques

Background:

  • Accurate assessment of antimicrobial activity in dental restorative materials requires reliable biofilm imaging.
  • Scanning electron microscopy (SEM) is crucial for visualizing biofilm structure but is sensitive to fixation methods.
  • Existing fixation protocols may not optimally preserve biofilm architecture for quantitative analysis.

Purpose of the Study:

  • To compare four biofilm fixation protocols for SEM imaging and Fiji analysis.
  • To introduce a complementary SEM cross-section analysis method for dental materials.
  • To validate the optimal fixation protocol with antimicrobial restorative materials.

Main Methods:

  • Resin composite discs incubated with Streptococcus mutans, followed by four fixation protocols (formalin/glutaraldehyde, ethanol dehydration).
  • SEM imaging and Fiji-based image analysis of biofilms and cross-sections.
  • Validation using resin composites and glass ionomer cement with antimicrobial agents.

Main Results:

  • Fixation protocols significantly impact SEM image quality, biofilm architecture, and quantitative analysis results.
  • Glutaraldehyde combined with graded ethanol dehydration yielded superior image resolution and discrimination.
  • The optimized protocol demonstrated improved sensitivity for detecting architectural differences in biofilm cross-sections.

Conclusions:

  • Standardized biofilm fixation is essential for reliable SEM analysis of dental materials.
  • Glutaraldehyde and ethanol dehydration provide superior biofilm preservation for SEM.
  • Machine-learning augmented analysis of SEM images enhances detection of biofilm structural changes.