Development of a high-throughput Candida albicans biofilm chip.
Development of a high-throughput Candida albicans biofilm chip.
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DOI：
10.1371/journal.pone.0019036
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发表时间：
2011-04-22
期刊：
影响因子：
3.7
通讯作者：
Ramasubramanian AK
中科院分区：
文献类型：
--
作者：
Srinivasan A;Uppuluri P;Lopez-Ribot J;Ramasubramanian AK
我们开发了一种由白色念珠菌纳米生物膜组成的高密度微阵列平台。使用机器人微点阵仪将封装在胶原蛋白基质中的白色念珠菌酵母细胞以低至 50 nL 的体积打印到表面修饰的显微镜载玻片上。孵化后，细胞生长成完全形成的“纳米生物膜”。通过扫描电子和共焦扫描激光显微镜评估这些生物膜的形态和结构复杂性。使用微阵列扫描仪根据 FUN 1 代谢过程引起的荧光强度变化来确定生物膜形成的程度。这种染色技术也适用于抗真菌药敏测试，结果表明，与常规生物膜类似，芯片上生物膜内的细胞对抗真菌药物（氟康唑和两性霉素 B）表现出较高水平的耐药性。因此，结构分析和抗真菌敏感性测试的结果表明，尽管微型化，这些生物膜仍显示出与生物膜生长模式相关的典型表型特性。在其最终格式中，白色念珠菌生物膜芯片 (CaBChip) 由单张载玻片上的 768 个等效且空间上不同的纳米生物膜组成；可以同时打印和处理多个芯片。与现有的微生物生物膜形成方法（即96孔微量滴定板模型）相比，这种真菌生物膜芯片具有小型化和自动化方面的优势，可减少试剂使用和分析时间，最大限度地减少劳动密集型步骤，并大幅降低检测成本。这种芯片能够同时快速、方便且廉价地筛选数百到数千种化合物，从而加速抗真菌药物的发现过程。
We have developed a high-density microarray platform consisting of nano-biofilms of Candida albicans. A robotic microarrayer was used to print yeast cells of C. albicans encapsulated in a collagen matrix at a volume as low as 50 nL onto surface-modified microscope slides. Upon incubation, the cells grow into fully formed “nano-biofilms”. The morphological and architectural complexity of these biofilms were evaluated by scanning electron and confocal scanning laser microscopy. The extent of biofilm formation was determined using a microarray scanner from changes in fluorescence intensities due to FUN 1 metabolic processing. This staining technique was also adapted for antifungal susceptibility testing, which demonstrated that, similar to regular biofilms, cells within the on-chip biofilms displayed elevated levels of resistance against antifungal agents (fluconazole and amphotericin B). Thus, results from structural analyses and antifungal susceptibility testing indicated that despite miniaturization, these biofilms display the typical phenotypic properties associated with the biofilm mode of growth. In its final format, the C. albicans biofilm chip (CaBChip) is composed of 768 equivalent and spatially distinct nano-biofilms on a single slide; multiple chips can be printed and processed simultaneously. Compared to current methods for the formation of microbial biofilms, namely the 96-well microtiter plate model, this fungal biofilm chip has advantages in terms of miniaturization and automation, which combine to cut reagent use and analysis time, minimize labor intensive steps, and dramatically reduce assay costs. Such a chip should accelerate the antifungal drug discovery process by enabling rapid, convenient and inexpensive screening of hundreds-to-thousands of compounds simultaneously.
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影响因子：
4.9
作者：
Nett, Jeniel E.;Crawford, Kyler;Andes, David R.
通讯作者：
Andes, David R.
影响因子：
11.8
作者：
Edmond, MB;Wallace, SE;Wenzel, RP
通讯作者：
Wenzel, RP
影响因子：
3.9
作者：
Garnier, G;Duskova-Smrckova, M;Revol, JF
通讯作者：
Revol, JF
DOI：
10.1086/651200
发表时间：
2010-07-01
期刊：
The Journal of infectious diseases
影响因子：
--
作者：
Nett JE;Sanchez H;Cain MT;Andes DR
通讯作者：
Andes DR
影响因子：
12.6
作者：
Fesenko, DO;Nasedkina, TV;Mirzabekov, AD
通讯作者：
Mirzabekov, AD