JAC Advance Access originally published online on March 20, 2006
Journal of Antimicrobial Chemotherapy 2006 57(6):1227-1230; doi:10.1093/jac/dkl096
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Induction of lethal photosensitization in biofilms using a confocal scanning laser as the excitation source
1 School of Dental Sciences, The University of Liverpool Liverpool, UK 2 Division of Microbial Diseases, UCL Eastman Dental Institute, University College London London, UK
Received 12 January 2006; returned 31 January 2006; revised 1 March 2006; accepted 2 March 2006
*Correspondence address. School of Dental Sciences, The University of Liverpool, Edwards Building, Daulby Street, Liverpool L69 3GN, UK. Tel: +44-151-706-5252; Fax: +44-151-706-5809; E-mail: c.hope{at}liv.ac.uk
Objectives: To induce lethal photosensitization in biofilms of Streptococcus pyogenes using the scanning laser in a confocal microscope to photoactivate Sn (IV) chlorin e6 (SnCe6) while simultaneously measuring changes in cell vitality using fluorescent indicators of membrane integrity.
Methods: Biofilms of S. pyogenes were immersed in a solution of 50 mg/L (70.28 µM) SnCe6 and scanned with the 488 nm argon and 543 nm HeNe lasers in a confocal microscope. Changes in membrane permeability were quantified using image analysis tools.
Results: Cell permeability increased in biofilms of S. pyogenes after successive scanning/exposure cycles in the presence of SnCe6.
Conclusions: Cell death was induced in biofilms of S. pyogenes by the photosensitizer SnCe6 on exposure to the scanning laser emissions of a confocal microscope. The simultaneous recording of cell death demonstrates the real-time evaluation of a light-activated antimicrobial compound against a biofilm.
Keywords: photodynamic therapy , Streptococcus pyogenes , Sn (IV) chlorin e6
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