Photobleaching of red fluorescence in oral biofilms

• Published in: Journal of periodontal research 2011-04, Vol.46 (2), p.228-234
• Main Authors: Hope, C. K., de Josselin de Jong, E., Field, M. R. T., Valappil, S. P., Higham, S. M.
• Format: Article
• Language: English
• Subjects: Bacteriological Techniques; Biofilms - radiation effects; Biological and medical sciences; Dental Plaque - microbiology; Dentistry; Filtration - instrumentation; Fluorescence; Humans; Image Processing, Computer-Assisted; Lighting - instrumentation; Medical sciences; oral biofilm; Otorhinolaryngology. Stomatology; Photobleaching; Photography - instrumentation; plaque; porphyrin; Porphyrins - radiation effects; Prevotella intermedia - radiation effects; Prevotella nigrescens - radiation effects; quantitative light-induced fluorescence digital; Radiation Dosage; Saliva - microbiology; Time Factors
• ISSN: 0022-3484; 1600-0765
• DOI: 10.1111/j.1600-0765.2010.01334.x

Hope CK, de Josselin de Jong E, Field MRT, Valappil SP, Higham SM. Photobleaching of red fluorescence in oral biofilms. J Periodont Res 2011; 46: 228–234. © 2010 John Wiley & Sons A/S Background and Objective:  Many species of oral bacteria can be induced to fluoresce due to the presence of endogenous porphyrins, a phenomenon that can be utilized to visualize and quantify dental plaque in the laboratory or clinical setting. However, an inevitable consequence of fluorescence is photobleaching, and the effects of this on longitudinal, quantitative analysis of dental plaque have yet to be ascertained. Material and Methods:  Filter membrane biofilms were grown from salivary inocula or single species (Prevotella nigrescens and Prevotella intermedia). The mature biofilms were then examined in a custom‐made lighting rig comprising 405 nm light‐emitting diodes capable of delivering 220 W/m2 at the sample, an appropriate filter and a digital camera; a set‐up analogous to quantitative light‐induced fluorescence digital. Longitudinal sets of images were captured and processed to assess the degradation in red fluorescence over time. Results:  Photobleaching was observed in all instances. The highest rates of photobleaching were observed immediately after initiation of illumination, specifically during the first minute. Relative rates of photobleaching during the first minute of exposure were 19.17, 13.72 and 3.43 arbitrary units/min for P. nigrescens biofilms, microcosm biofilm and P. intermedia biofilms, respectively. Conclusion:  Photobleaching could be problematic when making quantitative measurements of porphyrin fluorescence in situ. Reducing both light levels and exposure time, in combination with increased camera sensitivity, should be the default approach when undertaking analyses by quantitative light‐induced fluorescence digital.