Use of a High-throughput <em>In Vitro</em> Microfluidic System to Develop Oral Multi-species Biofilms

Derek S. Samarian; Nicholas S. Jakubovics; Ting L. Luo; Alexander H. Rickard

doi:10.3791/52467

Use of a High-throughput In Vitro Microfluidic System to Develop Oral Multi-species Biof...

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JoVE Journal Bioengineering

Use of a High-throughput In Vitro Microfluidic System to Develop Oral Multi-species Biofilms

Use of a High-throughput In Vitro Microfluidic System to Develop Oral Multi-species Biofilms

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07:09 min

December 1, 2014

DOI: 10.3791/52467-v

Derek S. Samarian¹, Nicholas S. Jakubovics², Ting L. Luo¹, Alexander H. Rickard¹

¹Department of Epidemiology, School of Public Health,The University of Michigan, ²Centre for Oral Health Research, School of Dental Sciences,Newcastle University

Overview

This methods paper describes a microfluidic system designed for the development of multi-species biofilms that mimic human supragingival dental plaque. The study emphasizes techniques for analyzing biofilm architecture, viability, and harvesting for further analyses.

Key Study Components

Area of Science

Microfluidics
Biofilm research
Dental microbiology

Background

Biofilms are complex communities of microorganisms.
Human dental plaque is a common example of a biofilm.
Understanding biofilm formation is crucial for dental health.
Current methods for studying biofilms have limitations.

Purpose of Study

To create representative multi-species dental plaque biofilms.
To develop a high-throughput system for biofilm analysis.
To improve methods for studying biofilm viability and architecture.

Methods Used

Collection of saliva samples from volunteers.
Preparation of representative media and inoculum.
Introduction of inoculum into a microfluidic chip.
Staining and imaging of biofilms using microscopy techniques.

Main Results

Successful formation of biofilms within microchannels.
High-throughput capability allows for multiple experiments.
Reduced material usage compared to traditional methods.
In situ imaging provides insights into biofilm structure.

Conclusions

The microfluidic system is an effective tool for biofilm research.
This method enhances the understanding of dental plaque biofilms.
Future studies can leverage this approach for various applications.

Frequently Asked Questions

What is the significance of studying biofilms?

Studying biofilms is crucial for understanding microbial interactions and their implications for health, particularly in dental contexts.

How does the microfluidic system improve biofilm research?

It allows for high-throughput experiments with reduced material usage, enabling more efficient studies of biofilm characteristics.

What techniques are used to analyze the biofilms?

Confocal laser scanning microscopy and epi fluorescence microscopy are used for in situ analysis of biofilm structure.

How are saliva samples collected for the study?

Saliva samples are collected from a cohort of volunteers in individual plastic tubes.

What are the advantages of this method over traditional flow cells?

The microfluidic system allows for parallel experiments and uses less material, making it more efficient than traditional flow cells.

Can this method be applied to other types of biofilms?

Yes, the microfluidic approach can be adapted for studying various types of biofilms beyond dental plaque.

The goal of this methods paper is to describe the use of a microfluidic system for the development of multi-species biofilms that contain species typically identified in human supragingival dental plaque. Methods to describe biofilm architecture, biofilm viability, and an approach to harvest biofilm for culture-dependent or culture-independent analyses are highlighted.

The overall aim of this procedure is to create compositionally representative, multi-species dental plaque biofilms. In parallel for analysis, this is achieved by first creating a representative media and inoculum. The inoculum is then introduced to the microfluidic chip after an overnight incubation, a biofilm forms inside the micro channel.

Finally, the biofilm is washed and stained for in situ two confocal laser scanning microscopy or epi fluorescence microscopy. The main advantage to this technique over existing methods like flow cells is that it's a high throughput system that allows for multiple experiments to be done in parallel while using less materials and not requiring artificial lab media. To begin collect saliva samples from a cohort of five or more volunteers in individual 50 milliliter plastic tubes.

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