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Related Concept Videos

Biofilms01:29

Biofilms

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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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Related Experiment Video

Updated: Mar 22, 2026

In vitro Biofilm Formation in an 8-well Chamber Slide
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Enterococcus Faecalis Biofilm. Formation and Development in Vitro Observed by Scanning Electron Microscopy.

María de Los Á Bulacio1, Lucas R Galván2, Cristina Gaudioso3

  • 1Department of Endodontics, School of Dentistry, National University of Tucumán, Tucumán, Argentina. maritabulacio@hotmail.com.

Acta Odontologica Latinoamericana : AOL
|April 21, 2016
PubMed
Summary

Formaldehyde fixation is the best method for detecting Enterococcus faecalis biofilms in root canals. Scanning electron microscopy confirmed biofilm development after 14 days of incubation.

Keywords:
Enterococcus faecalisbiofilmscanning electron microscopy

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Area of Science:

  • Microbiology
  • Dental Research
  • Biomaterials

Background:

  • Enterococcus faecalis is a common bacterium found in root canal infections.
  • Bacterial biofilms contribute to persistent infections and treatment failure.
  • Accurate detection of biofilm formation is crucial for understanding and managing endodontic infections.

Purpose of the Study:

  • To evaluate and compare different fixation methods for detecting Enterococcus faecalis biofilms on microplates.
  • To determine the optimal fixation technique for quantifying biofilm formation.
  • To correlate microplate-based detection with scanning electron microscopy (SEM) findings.

Main Methods:

  • Enterococcus faecalis biofilms were cultured on microplates and stained with crystal violet.
  • Three fixation methods were tested: no fixation, heat fixation, and 10% formaldehyde fixation.
  • Biofilm quantification was performed using a microplate reader.
  • Root portions were incubated with E. faecalis and analyzed using SEM at various time points.

Main Results:

  • 10% formaldehyde fixation yielded significantly higher biofilm detection compared to heat fixation or no fixation (ANOVA p<0.0001).
  • SEM analysis revealed consistent E. faecalis growth and confirmed biofilm development starting from 14 days of incubation.
  • Formaldehyde fixation proved to be the most effective method for detecting E. faecalis biofilms on microplates.

Conclusions:

  • 10% formaldehyde fixation is the superior technique for detecting Enterococcus faecalis biofilms in vitro.
  • SEM corroborates the microplate findings, indicating biofilm formation is established by 14 days.
  • This study provides a reliable method for assessing E. faecalis biofilm development in endodontic research.