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Method Article
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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No conflicting financial interests
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Posted 08 Mar, 2017
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Subject Areas
Microbiology
Method Article
Prashant Sharma
Biomedical Engineering UMCG
Corresponding Author
Version 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
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