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Use of a High-throughput In Vitro Microfluidic System to Develop Oral Multi-species Biofilms
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High-throughput quantitative method for assessing coaggregation among oral bacterial species.

E Levin-Sparenberg1, J M Shin2, E M Hastings3

  • 1Epidemiology Department, School of Public Health, University of Michigan, Ann Arbor, MI, USA.

Letters in Applied Microbiology
|July 26, 2016
PubMed
Summary
This summary is machine-generated.

Two new methods, a microplate assay and FlowCam(™) imaging, efficiently screen and quantify bacterial coaggregation. These tools aid in studying biofilm development by analyzing bacterial interactions.

Keywords:
ActinomycetesBiofilmsBiotechnologyCoaggregationStreptococci

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Area of Science:

  • Microbiology
  • Biofilm formation
  • Bacterial interactions

Background:

  • Coaggregation between bacterial species is crucial for the development of multi-species biofilms.
  • Previous limitations in quantifying coaggregation have hindered mechanistic studies.
  • Oral bacteria like Streptococcus gordonii, Streptococcus oralis, and Actinomyces oris exhibit well-characterized coaggregation patterns.

Purpose of the Study:

  • To develop and compare two quantitative methods for screening and analyzing bacterial coaggregation.
  • To assess the utility of a microplate reader and FlowCam(™) real-time imaging for coaggregation studies.
  • To provide tools for high-throughput screening and in-depth analysis of autoaggregation and coaggregation.

Main Methods:

  • A microplate-based assay was used to quantify cell density for coaggregation scoring 60 minutes postmixing.
  • High-speed real-time imaging with FlowCam(™) was employed to measure the rate of coaggregation over time.
  • Results from both methods were compared against visual coaggregation scores and microscopic observations using oral bacteria.

Main Results:

  • Both microplate and FlowCam(™) methods showed strong correlation with visual coaggregation scores.
  • The microplate assay proved effective for high-throughput screening of coaggregating bacterial pairs.
  • The FlowCam(™) assay provided detailed quantification of coaggregation rates and aggregate formation.

Conclusions:

  • Two complementary quantitative methods, a microplate assay and FlowCam(™) imaging, have been successfully demonstrated for bacterial coaggregation studies.
  • These assays overcome previous limitations in rapid and reproducible quantification, facilitating mechanistic investigations.
  • The developed methods enable both high-throughput screening and in-depth analysis, paving the way for future research on large bacterial panels.