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Method Article
A constant depth film fermenter to grow microbial biofilms
Prashant Sharma
Biomedical Engineering UMCG
Corresponding Author
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
This protocol describes how to grow biofilms with a well-defined thickness to match the thickness of clinically occurring biofilms in a constant-depth-film-fermenter. In a constant-depth-film-fermenter, biofilms are grown on the bottom of wells with set depths, while a scraper blade removes biofilm growing above the wells. Proper fixing of well-depth and use of smooth scraper blades are critical steps for growing biofilms of constant thickness over their entire surface area. Biofilm thickness can be measured with confocal-laser-scanning-microscopy, low-load-compression-testing or optical-coherence-tomography. Confocal-laser-scanning-microscopy is mostly used, but relies on penetration of fluorophores and laser-light through the biofilms. This makes confocal-laser-scanning-microscopy unsuitable for relatively thick constant-depth-film-fermenter biofilms, leaving low-load-compression-testing or optical-coherence-tomography preferred. Also, the relatively low resolution of optical-coherence-tomography enables to determine thickness over an entire biofilm surface area, constituting a major advantage over confocal-laser-scanning-microscopy. The reproducible thickness of constant-depth-film-fermenter biofilms facilitates high-throughput studies and is important for studying antimicrobial penetration in biofilms.
Keywords
biofilm, bacteria, fungi, infection, cystic fibrosis, burn wound
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This is a preprint. It has not completed peer review.
Method Article
A constant depth film fermenter to grow microbial biofilms
Prashant Sharma
Biomedical Engineering UMCG
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 08 Mar, 2017
No community comments so far
Metrics
Comments: 0
HTML Views: ...
Subject Areas
Microbiology
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
This protocol describes how to grow biofilms with a well-defined thickness to match the thickness of clinically occurring biofilms in a constant-depth-film-fermenter. In a constant-depth-film-fermenter, biofilms are grown on the bottom of wells with set depths, while a scraper blade removes biofilm growing above the wells. Proper fixing of well-depth and use of smooth scraper blades are critical steps for growing biofilms of constant thickness over their entire surface area. Biofilm thickness can be measured with confocal-laser-scanning-microscopy, low-load-compression-testing or optical-coherence-tomography. Confocal-laser-scanning-microscopy is mostly used, but relies on penetration of fluorophores and laser-light through the biofilms. This makes confocal-laser-scanning-microscopy unsuitable for relatively thick constant-depth-film-fermenter biofilms, leaving low-load-compression-testing or optical-coherence-tomography preferred. Also, the relatively low resolution of optical-coherence-tomography enables to determine thickness over an entire biofilm surface area, constituting a major advantage over confocal-laser-scanning-microscopy. The reproducible thickness of constant-depth-film-fermenter biofilms facilitates high-throughput studies and is important for studying antimicrobial penetration in biofilms.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6