Visualization of Biofilm Formation in Candida albicans Using an Automated Microfluidic Device.
Visualization of Biofilm Formation in Candida albicans Using an Automated Microfluidic Device.
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DOI：
10.3791/56743
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发表时间：
2017-12-14
期刊：
影响因子：
--
通讯作者：
Nobile CJ
中科院分区：
文献类型：
--
作者：
Gulati M;Ennis CL;Rodriguez DL;Nobile CJ
白色念珠菌是人类最常见的真菌病原体，约占医院获得性脓毒症病例的15%。C.白色念珠菌的另一个特征是其形成生物膜的能力，生物膜是附着在生物和非生物表面上的结构化细胞群落。C.白色念珠菌生物膜可在宿主组织（例如粘膜层）和医疗装置（例如导管、起搏器、假牙和关节假体）上形成。生物膜对临床提出了重大挑战，因为它们对物理和化学扰动具有高度抗性，并且可以作为种子传播感染的储存库。各种体外试验已被用于研究C。白念珠菌生物膜形成的测定，如微量滴定板测定、干重测量、细胞活力测定和共聚焦扫描激光显微镜。所有这些测定都是单终点测定，其中在特定时间点评估生物膜形成。在这里，我们描述了一个协议，研究生物膜的形成，在层流条件下使用自动化微流体装置实时。该方法允许观察生物膜形成，因为生物膜随着时间的推移而发展，使用模拟宿主的那些的可定制条件，例如在血管导管中遇到的那些。该方案可用于评估生物膜缺陷的遗传突变体，以及抗菌剂对生物膜发展的抑制作用，在实时。
Candida albicans is the most common fungal pathogen of humans, causing about 15% of hospital-acquired sepsis cases. A major virulence attribute of C. albicans is its ability to form biofilms, structured communities of cells attached to biotic and abiotic surfaces. C. albicans biofilms can form on host tissues, such as mucosal layers, and on medical devices, such as catheters, pacemakers, dentures, and joint prostheses. Biofilms pose significant clinical challenges because they are highly resistant to physical and chemical perturbations, and can act as reservoirs to seed disseminated infections. Various in vitro assays have been utilized to study C. albicans biofilm formation, such as microtiter plate assays, dry weight measurements, cell viability assays, and confocal scanning laser microscopy. All of these assays are single end-point assays, where biofilm formation is assessed at a specific time point. Here, we describe a protocol to study biofilm formation in real-time using an automated microfluidic device under laminar flow conditions. This method allows for the observation of biofilm formation as the biofilm develops over time, using customizable conditions that mimic those of the host, such as those encountered in vascular catheters. This protocol can be used to assess the biofilm defects of genetic mutants as well as the inhibitory effects of antimicrobial agents on biofilm development in real-time.
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影响因子：
6.7
作者：
Uppuluri P;Chaturvedi AK;Srinivasan A;Banerjee M;Ramasubramaniam AK;Köhler JR;Kadosh D;Lopez-Ribot JL
通讯作者：
Lopez-Ribot JL
影响因子：
64.5
作者：
Nobile CJ;Fox EP;Nett JE;Sorrells TR;Mitrovich QM;Hernday AD;Tuch BB;Andes DR;Johnson AD
通讯作者：
Johnson AD
影响因子：
9.2
作者：
Nobile, CJ;Mitchell, AP
通讯作者：
Mitchell, AP
影响因子：
3.1
作者：
Andes, D;Nett, J;Pitula, A
通讯作者：
Pitula, A
影响因子：
3.1
作者：
Nett, Jeniel E.;Marchillo, Karen;Andes, David R.
通讯作者：
Andes, David R.