ArticleApplied and environmental microbiology2010
Diffusion measurements inside biofilms by image-based fluorescence recovery after photobleaching (FRAP) analysis with a commercial confocal laser scanning microscope.
Article in Applied and environmental microbiology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed.
- Biofilm Analysis by Confocal Microscopy-Basics and Practical Aspects.Microscopy research and technique · 2026Review
- Beneficial applications of biofilms.Nature reviews. Microbiology · 2024Review
- Single Microcolony Diffusion Analysis in Pseudomonas aeruginosa Biofilms.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Altering the viscoelastic properties of mucus-grownBiofilm · 2023Article
- What's past is prologue: FRAP keeps delivering 50 years later.Biophysical journal · 2023Review
- Article
- Antimicrobial Photodynamic Inactivation Using Topical and Superhydrophobic Sensitizer Techniques: A Perspective from Diffusion in Biofilms.Photochemistry and photobiology · 2021Review
- Shear stress affects the architecture and cohesion of Chlorella vulgaris biofilms.Scientific reports · 2021Article
- Quantifying the effects of antibiotic treatment on the extracellular polymer network of antimicrobial resistant and sensitive biofilms using multiple particle tracking.NPJ biofilms and microbiomes · 2021Article
- Improved FRAP Measurements on Biofilms.Biophysical journal · 2020Article
- Microbial Extracellular Polymeric Substances (EPSs) in Ocean Systems.Frontiers in microbiology · 2017Review
- Article
- Lipid-Bilayer Dynamics Probed by a Carbon Dot-Phospholipid Conjugate.Biophysical journal · 2016Article
- Material properties of biofilms-a review of methods for understanding permeability and mechanics.Reports on progress in physics. Physical Society (Great Britain) · 2015Review
- Patterned hydrophobic domains in the exopolymer matrix of Shewanella oneidensis MR-1 biofilms.Applied and environmental microbiology · 2013Article
- Spatiotemporal dynamics of complement C5a production within bacterial extracellular polymeric substance.Journal of innate immunity · 2013Article
- Simplified equation to extract diffusion coefficients from confocal FRAP data.Traffic (Copenhagen, Denmark) · 2012Article
- Correlative time-resolved fluorescence microscopy to assess antibiotic diffusion-reaction in biofilms.Antimicrobial agents and chemotherapy · 2012Article
- Stenotrophomonas maltophilia: an emerging global opportunistic pathogen.Clinical microbiology reviews · 2012Review
- Single microcolony diffusion analysis inNPJ biofilms and microbiomesArticle
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Research about the reactional and structural dynamics of biofilms at the molecular level has made great strides, owing to efficient fluorescence imaging methods in terms of spatial resolution and fast acquisition time but also to noninvasive conditions of observation consistent with in situ biofilm studies. In addition to conventional fluorescence intensity imaging, the fluorescence recovery after photobleaching (FRAP) module can now be routinely implemented on commercial confocal laser scanning microscopes (CLSMs). This method allows measuring of local diffusion coefficients in biofilms and could become an alternative to fluorescence correlation spectroscopy (FCS). We present here an image-based FRAP protocol to improve the accuracy of FRAP measurements inside "live" biofilms and the corresponding analysis. An original kymogram representation allows control of the absence of perturbing bacterial movement during image acquisition. FRAP data analysis takes into account molecular diffusion during the bleach phase and uses the image information to extract molecular diffusion coefficients. The fluorescence spatial intensity profile analysis used here for the first time with biofilms is supported both by our own mathematical model and by a previously published one. This approach was validated to FRAP experiments on fluorescent-dextran diffusion inside Lactococcus lactis and Stenotrophomonas maltophilia biofilms, and the results were compared to previously published FCS measurements.
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