The Analysis Of Bacterial Biofilm From Chronic Refractory Wound In Vitro Model

Objective:To establish chronic refractory wound clinical common bacteria biofilm in vitro model by using laser confocal microscopy bacterial biofilm formation process, for further research on biological mechanism of membrane and chronic wound healingMethods:The bacterial biofilm in vitro model was established to compare the traditional first flat culture method and the guidance of culture method in order to guide culture biological membrane method in vitro, observe different time points and temperature on the influence of the biological membrane formation. Laser confocal microscope was used to the observe the formation processes of biofilm each strains. The FITC-ConA and PI respectively polysaccharide and mark bacterial DNA, the confocal laser scanning microscopy observation biofilm formation.Results:The traditional flat culture method finally was obseved the result with poor stability, and guide piece of culture method finally results more stablability, test a better repeatability,37℃of membrane is better than22℃in growth state. Each strains are scattered green fluorescent, and not dense by training after24hours. Green fluorescence significantly increased, and part and red fluorescent overlap, formation in the field of yellow fluorescence, staphylococcus aureus is obvious after48hours. The green fluorescent intensity and distribution, the strains of the clinical are more evident than standard strains. Pseudomonas aeruginosa and its standard strains of the green fluorescent increase, when the training lasts48hours. Green fluorescent is scattered, when the training lasts5days. Staphylococcus aureus in48hours of biofilm formation mature, pseudomonas aeruginosa is in72hours. Conclusions:The biological membrane of staphylococcus aureus and pseudomonas aeruginosa are common to produce from chronic wounds in clinical;By immune fluorescence technology and the confocal laser scanning microscopy research on biological film forming process are simple method. The formation of staphylococcus aureus and pseudomonas aeruginosa membrane time are different, biological film formation and characteristics have significant differences.