本文選題：白色念珠菌 + 生物被膜�。� 參考：《重慶醫(yī)科大學(xué)》2010年碩士論文

【摘要】： 第一部分 【目的】建立白色念珠菌(Candida albicans)生物被膜(Biofilm,BF)體外模型,運(yùn)用多種形態(tài)學(xué)方法檢測(cè),初步探討白色念珠菌BF相關(guān)感染發(fā)生機(jī)制。 【方法】體外建立2、8、24,48及72 h時(shí)間組白色念珠菌BF模型,通過銀染、熒光探針FITC-conA和SYTO9/PI標(biāo)記等不同途徑,結(jié)合熒光顯微鏡、掃描電鏡(scanning electron microscope, SEM),激光共聚焦顯微鏡(Confocal Laser Scanning Microscopy,CLSM)觀察不同時(shí)期白色念珠菌BF形成情況。 【結(jié)果】白色念珠菌能夠在玻片上形成典型的BF,通過熒光顯微鏡,SEM和CLSM完成對(duì)白色念珠菌BF形成過程動(dòng)態(tài)觀察,白色念珠菌隨著時(shí)間增加逐漸增加,48 h形成初步生物被膜,72 h結(jié)構(gòu)更加復(fù)雜和成熟。 【結(jié)論】平板法能夠較好地建立白色念珠菌BF體外模型,并可利用多種方式檢測(cè)。 第二部分 【目的】應(yīng)用ISA軟件及Image-Pro Plus 6.0軟件定量化分析白色念珠菌BF形成發(fā)展過程中變化,為進(jìn)一步深研究其耐藥機(jī)制及干預(yù)提供基礎(chǔ)。 【方法】將結(jié)合熒光探針SYTO9/PI標(biāo)記的CLSM圖片分別導(dǎo)入ISA軟件及Image-Pro Plus 6.0軟件分析獲得白色念珠菌BF相關(guān)結(jié)構(gòu)參數(shù)定量化數(shù)據(jù)。 【結(jié)果】①ISA軟件定量化分析顯示,SYTO9/PI熒光探針標(biāo)記的生物被膜模型的區(qū)域孔徑(AP)在24, 48和72 h分別為0.95±0.06, 0.89±0.01和0.83±0.01,平均擴(kuò)散距離(ADD)分別為1.16±0.13, 1.26±0.06和2.43±0.76,結(jié)構(gòu)熵(TE)分別為4.87±0.34, 5.18±0.35和5.47±0.16,兩兩比較,差異均有顯著性(P0.05)。②Image-Pro Plus 6.0軟件分析顯示,生物被膜內(nèi)真菌死亡率分別為(34.71±2.72)%, (36.63±4.20)%和( 47.41±2.53)%,與24 h及48 h相比,72 h真菌死亡率增加具有顯著性(P0.05)。 【結(jié)論】利用ISA軟件及Image-Pro Plus 6.0軟件定量化分析可以反映白色念珠菌BF形成過程中的空間結(jié)構(gòu)及死活菌比例變化特征。
[Abstract]:Part one [objective] to establish Candida albicansa biofilm biofilm BFs model in vitro and to investigate the mechanism of BF- related infection of Candida albicansa using various morphological methods. [methods] the BF model of Candida albicans was established in vitro by silver staining, fluorescence probe FITC-conA and SYTO9/PI labeling. Scanning electron microscopy (SEM) and confocal Laser Scanning microscopy (CLSM) were used to observe the formation of Candida albicans in different periods. [results] Candida albicans were able to form typical BFs on glass slides. The formation process of BF was observed by fluorescence microscope and CLSM. The structure of Candida albicans was more complex and mature at 72 h after 48 h of initial biofilm formation. [conclusion] the in vitro model of Candida albicans BF can be established by plate method, and it can be detected in many ways. Part two [objective] to analyze quantitatively the changes in the formation and development of Candida albicans BF by ISA software and Image-Pro Plus 6.0 software, so as to provide a basis for further study on the mechanism of drug resistance and intervention. [methods] the CLSM images labeled with fluorescent probe SYTO9/PI were imported into ISA software and Image-Pro Plus 6.0 software respectively to obtain quantitative data of BF related structure parameters of Candida albicans. [results] the quantitative analysis of 1ISA software showed that the APs of the biofilm model labeled with fluorescent probes of SYTO9 / Pi were 0.95 鹵0.06,0.89 鹵0.01,0.83 鹵0.01 at 24,48 and 72 h, respectively, and the average diffusion distances were 1.16 鹵0.13,1.26 鹵0.06 and 2.43 鹵0.76, respectively, and the structure entropy was 4.87 鹵0.34, respectively, and the mean diffusion distance was 1.16 鹵0.13,1.26 鹵0.06 and 2.43 鹵0.76, respectively, and the average diffusion distance was 1.16 鹵0.13,1.26 鹵0.06 and 2.43 鹵0.76, respectively. 5.18 鹵0.35 and 5.47 鹵0.16, respectively. The results showed that the mortality rates of biofilm fungi were 34.71 鹵2.72%, 36.63 鹵4.20% and (47.41 鹵2.53)%, respectively. The mortality of fungi in biofilm was significantly higher than that in 24 h and 48 h. [conclusion] the quantitative analysis of ISA and Image-Pro Plus 6.0 software can reflect the spatial structure of Candida albicans during the formation of BF and the variation characteristics of the proportion of live and dead bacteria.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R379.4

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本文編號(hào)：1796106