Establishment And Analysis Of The Model Of Candida Albicans Biofilm In Vitro

PartⅠ【Objective】To establish Candida albicans biofilm in vitro and discuss the mechanisms of infections caused by Candida albicans.【Methods】Candida albicans biofilm model in vitro was constructed on glass slice and its development was monitored at different time intervals (2hours,8 hours, 24 hours, 48hours, 72 hours). The biofilm was stained by silver, FITC-conA and SYTO9/PI, espectively. Ivestigated the biofilm forming process by fluorescence microscope, canning electron microscope (SEM) and confocal laser scanning microcope (CLSM).【Results】The model of Candida albicans biofilm in vitro could established on coverslip. Observation showed that biofilm formation was observed about 48 h after culture, and its architecture became more mature and complex after 72 h.【Conclusions】Candida albicans has biofilm-forming abilities, the biofilm can be identified and analyzed by many methods. PartⅡ【Objective】To quantify the development process of Candida albicans biofilm spatial structure with Image Structure Analyer (ISA) software and Image-Pro Plus 6.0 software. To provide assist to further investigation for the mechanism of drug resistance and the treatment of biofilm-associated infections.【Methods】The fluorescence images stack of different layers in Candida albicans biofilm model was obtained by CLSM, based on nucleic acid SYTO9/PI stained. Quantitative parameters of Candida albicans biofilm spatial structure could be acquired after the image information was calculated by ISA software and Image-Pro Plus 6.0 software.【Results】①The quantitative data from ISA software showed that in the biofilm models stained with SYTO9/PI at different time intervals(24 h, 48 h, 72 h), the areal porosity (AP) were 0.95±0.06, 0.89±0.01 and 0.83±0.01, respectively, the average diffusion distance (ADD) were 1.16±0.13, 1.26±0.06 and 2.43±0.76, respectively, the textural entropy (TE) were 4.87±0.34, 5.18±0.35 and 5.47±0.16, respectively. There were significantly differences between different two groups(P<0.05).②The quantitative data from Image-Pro Plus 6.0 software showed that the mortality of Candida albicans were (34.71±2.72)%, (36.63±4.20)% and ( 47.41±2.53)%, respectively. There were significantly differences between group72 h and other two groups (P<0.05).【Conclusions】The ISA software and Image-Pro Plus 6.0 software can provide useful information of Candida albicans biofilm spatial structure and the change of live-dead rate in biofilm process.