Diffusion measurements inside biofilms by image-based fluorescence recovery after photobleaching (FRAP) analysis with a commercial confocal laser scanning microscope.

Applied and Environmental Microbiology
François WaharteM P Fontaine-Aupart

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 a...Continue Reading

References

Jan 1, 1991·Biosensors & Bioelectronics·C C Cronenberg, J C van den Heuvel
Dec 15, 2004·Biophysical Journal·François WaharteFrançois Amblard
Jan 28, 2005·Antimicrobial Agents and Chemotherapy·Suriani Abdul RaniPhilip S Stewart
Nov 30, 2005·Comptes rendus biologies·Pascaline Lacroix-GueuMarie-Pierre Fontaine-Aupart
Apr 1, 1994·Applied and Environmental Microbiology·J R LawrenceD R Korber
Jun 1, 2007·Canadian Journal of Microbiology·J R LawrenceT R Neu
Aug 7, 2007·Journal of Bacteriology·Hans-Curt FlemmingDaniel J Wozniak
Feb 5, 2008·Applied and Environmental Microbiology·R BriandetM-P Fontaine-Aupart
Jan 27, 2009·Applied and Environmental Microbiology·Shoji TakenakaPhilip S Stewart
Mar 28, 2009·PLoS Pathogens·Luyan MaDaniel J Wozniak

❮ Previous
Next ❯

Citations

Mar 28, 2012·Antimicrobial Agents and Chemotherapy·S Daddi OubekkaK Steenkeste
Dec 12, 2012·Applied and Environmental Microbiology·Fadi AldeekJean-Claude Block
Jan 11, 2012·Clinical Microbiology Reviews·Joanna S Brooke
Apr 6, 2013·International Journal of Food Microbiology·J FlouryS Lortal
Aug 16, 2014·Journal of Controlled Release : Official Journal of the Controlled Release Society·Katrien ForierKevin Braeckmans
Sep 19, 2012·Traffic·Minchul KangEmmanuele DiBenedetto
Nov 30, 2011·Biotechnology and Bioengineering·A S Van WeyP R Shorten
Mar 17, 2016·Journal of Microbiological Methods·Sebastian Schlafer, Rikke L Meyer
Jan 28, 2015·Trends in Microbiology·Thomas R Neu, John R Lawrence
May 12, 2016·Biophysical Journal·Sukhendu NandiRaz Jelinek
Jul 1, 2017·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Rym BoudjemaaKarine Steenkeste
May 3, 2018·FEMS Microbiology Letters·Jagadish SankaranThorsten Wohland
Oct 30, 2019·Expert Review of Anti-infective Therapy·Samantha Flores-TreviñoElvira Garza-González
Feb 27, 2015·Reports on Progress in Physics·Nicole BillingsKatharina Ribbeck
Nov 16, 2019·NPJ Biofilms and Microbiomes·Jagadish SankaranThorsten Wohland
Jun 14, 2017·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Senthilguru KulanthaivelIndranil Banerjee
Jun 13, 2017·Frontiers in Microbiology·Alan W Decho, Tony Gutierrez
Nov 8, 2020·NPJ Biofilms and Microbiomes·Jagadish SankaranThorsten Wohland
Mar 2, 2021·Journal of Molecular Biology·Debabrata DeyGideon Schreiber
Apr 19, 2020·Biophysical Journal·Jan HauthRoland Ulber
Jun 6, 2021·International Journal of Pharmaceutics·Li DingZhengrong Cui
Mar 15, 2012·Langmuir : the ACS Journal of Surfaces and Colloids·Nadia BourouinaNelly Henry
Aug 20, 2021·Critical Reviews in Microbiology·Kecheng QuanHenny C van der Mei
Jan 6, 2018·Journal of Agricultural and Food Chemistry·Xiu Hang ChaiYuan Fa Liu

❮ Previous
Next ❯

Related Concepts

Related Feeds

Biofilm & Infectious Disease

Biofilm formation is a key virulence factor for a wide range of microorganisms that cause chronic infections.Here is the latest research on biofilm and infectious diseases.