Study on Method for Observation of Bacterial Biofilm Formation under Confocal Laser Scanning Microscope

Objective:To establish the method of observation of bacterial biofilm formation under confocal laser scanning microscope(CLSM),so as to provide the basis for further study on the mechanism of biofilm formation.Methods:X428,a clinical isolation of Staphylococcus Aureus(SA),was selected to be investigated.Cover glasses were used to culture biofilms and collected at 4th,8th,12th,16th,24th and 48th hour of incubation.Polysaccharides and bacteria were labeled by immunofluorescent technique. The condition of SA biofilm formation was observed under confocal laser scanning microscope(CLSM).Results:A series of images of biofilm were collected by CLSM.Tiny colonies adhered on glasses at the 4th hour of incubation.Bacteria aggregated into dusters and polysaccharides were created at the 8th and 12th hour.Polysaccharides accumulated gradually until mature biofilms were formed at the 16th hour.Biofilms had dispersed at the 24th and 48th hour.Conclusions:It's a simple and feasible method to study the process of biofilm formation by using immunofluorescent technique and confocal laser scanning microscope.