DETECTION OF BIOFILMS ON ADENOIDS AND CHOLESTEATOMAS USING CONFOCAL SCANNING LASER MICROSCOPY
M.H. Ng*, Y.S. Ee, A.J. Sow, and B.S. Goh
Abstract
DETECTION OF BIOFILMS ON ADENOIDS AND CHOLESTEATOMAS USING CONFOCAL SCANNING LASER MICROSCOPYM.H. Ng1*, Y.S. Ee2, A.J. Sow2, and B.S. Goh2
1Tissue Engineering Centre, Univerisiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, 56000 Kuala Lumpur, MALAYSIA.
2 Department of Otorhinolaryngology, Univerisiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, 56000 Kuala Lumpur, MALAYSIA.
ABSTRACT
Biofilm is an organized bacterial community embedded in extracellular polymeric matrix that has been associated with periodic occurrence and chronic infection in certain diseases. Biofilm often escape routine culture screening but recent advances in microscopy and molecular technologies have made it possible to examine biofilm in detail. With confocal scanning laser microscopy (CSLM), it is possible to study the three-dimensional structure of the biofilm aided with computer- enhanced microscopy analyses and 3D computer reconstruction. In this study, presence of biofilms on adenoids and cholesteatomas, both presents as a recurrent infection problem in otorhinolaryngology were investigated. Tissues were obtained with prior written consent from patients in UKM Medical Centre. The specimens were first snap-frozen in liquid nitrogen. Using cryo-microtomy, tissue sections of 5 micron were prepared on glass slides. The sections were stained with Propidium Iodide (PI) to stain bacterial cells nuclei red then incubated with concanavalin A (Con A) fluorescein isothiocyanate (FITC) to stain the glycocalyx matrix green. The sections were then examined using CLSM. We successfully detected the presence of biofilms in 27/49 (55.1%) adenoid tissues and 15/27 (55.5%) cholesteatomas. In conclusion, biofilms do persist on adenoid and choleasteatoma tissues and CLSM may be a reliable technique to detect biofilms.
Keywords: CSLM, middle ear disease, mastoid, bacterial colony, ear infection
