Name: candida albicans biofilm chip (cabchip) for high-throughput antifungal drug screening
Uploaded: 2012-07-18T11:00:00
Description: Watch this Scientific Journal Video about Candida albicans Biofilm Chip CaBChip for High-throughput Antifungal Drug Screening at JoVE.com

这项工作的部分资金由来自南得克萨斯科技管理（POCrr 2009.041），整合从国家研究资源中心（UL 1RR025767）医学和科学学院的赠款和牙科及颅面研究国家研究所（ 5R21DE017294）。

1。制备官能幻灯片<ol><li>载玻片放置在一个可移动的滑动机架，洗浸泡在罐子染色，含99％乙醇（病理分级）的两倍。用幻灯片使用纸巾（确保不产生纸尘），干用压缩氮气射流清洗。 </li></ol> 注意：不要使用金的湿巾擦拭幻灯片，因为它会产生细尘纸。 <ol start="2"><li>沉浸充满在室温下与浓硫酸和孵育过夜染色罐中含有幻灯片幻灯片架。 </li></ol><p class="jove_step"...

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	<li>Edmond, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nosocomial+bloodstream+infections+in+United+States+hospitals:+a+three-year+analysis.">Nosocomial bloodstream infections in United States hospitals: a three-year analysis.</a> Clin. Infect. Dis. 29, 239-244 (1999).</li><li>Ramage, G., Bachmann, S., Patterson, T. F., Wickes, B. L., Lopez-Ribot, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigation+of+multidrug+efflux+pumps+in+relation+to+fluconazole+resistance+in+Candida+albicans+biofilms.">Investigation of multidrug efflux pumps in relation to fluconazole resistance in Candida albicans biofilms.</a> J. Antimicrob. Chemother. 49, 973-980 (2002).</li><li>Srinivasan, A., Uppuluri, P., Lopez-Ribot, J., Ramasubramanian, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+High-Throughput+Candida+albicans+Biofilm+Chip.">Development of a High-Throughput Candida albicans Biofilm Chip.</a> PLoS ONE. 6, 19036-19036 (2011).</li><li>Ramage, G., Vandewalle, K., Wickes, B. L., Lopez-Ribot, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characteristics+of+biofilm+formation+by+Candida+albicans.">Characteristics of biofilm formation by Candida albicans.</a> Rev. Iberoam. Micol. 18, 163-170 (2001).</li><li>Pierce, C. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+and+reproducible+96-well+plate-based+method+for+the+formation+of+fungal+biofilms+and+its+application+to+antifungal+susceptibility+testing.">A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing.</a> Nat. Protoc. 3, 1494-1500 (2008).</li><li>Lee, M. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+cellular+microarray+for+high-throughput+toxicology+assays.">Three-dimensional cellular microarray for high-throughput toxicology assays.</a> Proc. Natl. Acad. Sci. U.S.A. 105, 59-63 (2008).</li><li>Meyerhofer, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characteristics+of+resist+films+produced+by+spinning.">Characteristics of resist films produced by spinning.</a> Journal of Applied Physics. 49, (1978).</li><li>Ramage, G., Vande Walle, K., Wickes, B. L., Lopez-Ribot, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Standardized+method+for+in+vitro+antifungal+susceptibility+testing+of+Candida+albicans+biofilms.">Standardized method for in vitro antifungal susceptibility testing of Candida albicans biofilms.</a> Antimicrob. Agents Chemother. 45, 2475-2479 (2001).</li><li>Chandra, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biofilm+formation+by+the+fungal+pathogen+Candida+albicans:+Development,+architecture,+and+drug+resistance.">Biofilm formation by the fungal pathogen Candida albicans: Development, architecture, and drug resistance.</a> Journal of Bacteriology. 183, 5385-5394 (2001).</li><li>Jabra-Rizk, M. A., Falkler, W. A., Meiller, T. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fungal+biofilms+and+drug+resistance.">Fungal biofilms and drug resistance.</a> Emerg. Infect. Dis. 10, 14-19 (2004).</li><li>Tobudic, S., Lassnigg, A., Kratzer, C., Graninger, W., Presterl, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antifungal+activity+of+amphotericin+B,+caspofungin+and+posaconazole+on+Candida+albicans+biofilms+in+intermediate+and+mature+development+phases.">Antifungal activity of amphotericin B, caspofungin and posaconazole on Candida albicans biofilms in intermediate and mature development phases.</a> Mycoses. 53, 208-214 (2010).</li></ol>

我们已经开发了一个基于细胞的高密度芯片，CA BChip， 白念珠菌生物膜的纳升量组成。芯片上印改性玻璃基板，允许强大的胶原蛋白凝胶点附件，同时提供必要非蔓延，半球形的三维凝胶疏水。一个单一的钙 BChip可以取代约八96孔板，和几个芯片可以打印，并在同一时间处理。该芯片采用纳米级的文化，可以方便，快捷的处理，适合自动化，最大限度地减少手工劳动和成本，?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td> 3 - 氨丙基（APTES） </td><td> Sigma-Aldrich公司</td><td> 440140 </td></tr><tr><td>聚苯乙烯 - 马来酸酐共聚物（PS-MA的） </td><td> Sigma-Aldrich公司</td><td> 426946 </td></tr><tr><td>玻璃显微镜幻灯片</td><td> Fisher Scientific则</td><td> 12-549-3 </td></tr><tr><td>鼠尾胶原蛋白I型</td><td> BD公司</td><td> 354236 </td></tr><tr><td>机器人微阵列</td><td> omn​​igrid微</td><td> MICROSYS4000/4100A </td></tr><tr><td>微阵列扫描仪</td><td> genepix个人4100A </td><td> GENEPIX4100A </td></tr><tr><td>杂交盒式</td><td> ArrayIt公司</td><td> AHCXD </td></tr><tr><td> FUN1 [2-chloRO-4-（2,3 - 二氢-3 - 甲基 - （苯并-1,3 - 噻唑-2 - 基） - 亚甲基）-1 - 苯基quinoliniumiodide] </td><td> Invitrogen公司</td><td>架F-7030 </td></tr><tr><td>氟康唑</td><td> sicor制药公司</td><td> J02AC01 </td></tr><tr><td>两性霉素B </td><td>西格玛</td><td> A2411 </td></tr><tr><td>的RPMI-1640 </td><td>敏达，公司</td><td> 50-020-PC </td></tr><tr><td>陶瓷提示190微米的小孔</td><td> digilab </td><td> 60020441-00 </td></tr><tr><td> GraphPad棱镜软件</td><td> GraphPad软件公司</td><td></td></tr><tr><td> genepix临V4.1的</td><td>分子器件</td><td></td></tr></tbody></table>

candida albicans biofilm chip (cabchip) for high-throughput antifungal drug screening

白念珠菌仍然是主要病原念珠菌，目前第四个最常见的在美国的医院院内血流感染。这些机会性感染造成损害个人越来越多越来越大的威胁，并进行不可接受的高死亡率。这是由于抗真菌药物的有限阿森纳的一部分，但也出现抵抗最常用的抗真菌药物。进一步复杂化的治疗方法是，多数表现念珠菌生物膜的形成与相关的事实，并在这些生物膜的细胞显示临床使用抗真菌药耐药水平的提高。在这里，我们描述的C.组成的高密度芯片的发展白念珠菌生物膜纳米，我们有一个名为钙 BChip 3。简言之，一个机器人微阵列用tØ打印C.酵母细胞到固体表面的白色念珠菌 。在印刷过程中，酵母细胞被包围在一个三维矩阵使用量低至50 NL，并固定在一个合适的涂层的玻璃基板。经过最初的印刷，幻灯片培养24小时，使生物膜的发展，在37°C。在此期间的景点成长为全面发展的“纳米生物膜”，显示典型的结构和表型特征与成熟的三白念珠菌生物膜（ 即形态的复杂性，三维结构和耐药性）4。总体而言， 钙 BChip是〜750相当于和空间不同生物膜的组成与打印多个芯片可以同时处理额外的好处。估计FUN1代谢的荧光强度染色用芯片扫描仪通过测量细胞活力。这种真菌芯片非常适合为u在真正的高通量筛选抗真菌药物的发现本身。与现行标准（ 即 96酶标板模型生物膜形成5）相比，真菌生物膜芯片的主要优点是自动化，小型化，节省试剂和分析时间的量和成本，以及消除劳动密集的步骤。我们相信，这种芯片将显着加快的抗真菌的药物发现过程。

Watch this Scientific Journal Video about Candida albicans Biofilm Chip CaBChip for High-throughput Antifungal Drug Screening at JoVE.com

Candida albicans Biofilm Chip CaBChip for High-throughput Antifungal Drug Screening

我们已经开发出一种高密度芯片平台的3D纳米生物膜的组成<em&gt; C。白念珠菌</em&gt;称为<em&gt;钙</em&gt; BChip。上测试药物的敏感性个人资料<em&gt;钙</em&gt; BChip是传统的96孔板模式相媲美，表明真菌的芯片非常适合真正的高通量筛选抗真菌药物。

白色念珠菌生物膜芯片（CA BChip）为高通量的抗真菌药物筛选

Candida  albicans remains the main etiological agent of candidiasis, which  currently represents the fourth most common nosocomial bloodstream infection ...

Research

JoVE Journal

Bioengineering

Candida albicans Biofilm Chip (CaBChip) for High-throughput Antifungal Drug Screening

Cell-based Calcium Assay for Medium to High Throughput Screening of TRP Channel Functions using FlexStation 3

基于细胞中钙含量TRP通道功能的高通量筛选，使用FlexStation 3

The Molecular Devices' FlexStation 3 is a benchtop multi-mode microplate reader capable of automated fluorescence measurement in multi-well plates.  It is ...

科研

生物工程

Polymer Microarrays for High Throughput Discovery of Biomaterials

聚合物芯片高通量发现的生物材料

The discovery of novel biomaterials that are optimized for a specific biological application is readily achieved using polymer microarrays, which allows a ...

A Rapid and Quantitative Fluorimetric Method for Protein-Targeting Small Molecule Drug Screening

一种快速定量荧光方法蛋白靶向小分子药物筛选

We demonstrate a new drug screening method for determining the binding affinity of small drug molecules to a target protein by forming fluorescent gold ...

Using Tomoauto: A Protocol for High-throughput Automated Cryo-electron Tomography

使用Tomoauto：一个协议，用于高通量自动化低温电子断层扫描

Cryo-electron tomography (Cryo-ET) is a powerful three-dimensional (3-D) imaging technique for visualizing macromolecular complexes in their native ...

A Microfluidic Platform for High-throughput Single-cell Isolation and Culture

高通量单细胞的分离和文化的微流体平台

Studying the heterogeneity of single cells is crucial for many biological questions, but is technically difficult. Thus, there is a need for a simple, yet ...

High-throughput Identification of Bacteria Repellent Polymers for Medical Devices

医疗器械细菌高通量鉴定驱蚊聚合物

Medical devices are often associated with hospital-acquired infections, which place enormous strain on patients and the healthcare system as well as ...

Automated Contraction Analysis of Human Engineered Heart Tissue for Cardiac Drug Safety Screening

人工程心脏组织的收缩自动分析心脏药物安全筛选

Cardiac tissue engineering describes techniques to constitute three dimensional force-generating engineered tissues. For the implementation of these ...

3D Microtissues for Injectable Regenerative Therapy and High-throughput Drug Screening

3D Microtissues 用于可注射再生疗法和高通量药物筛选

To upgrade traditional 2D cell culture to 3D cell culture, we have integrated microfabrication with cryogelation technology to produce macroporous ...

Use of High-Throughput Automated Microbioreactor System for Production of Model IgG1 in CHO Cells

高通量自动 Microbioreactor 系统在 CHO 细胞模型 IgG1 生产中的应用

Automated microscale bioreactors (15 mL) can be a useful tool for cell culture engineers. They facilitate the simultaneous execution of a wide variety of ...

Applications for Open Source Microplate-Compatible Illumination Panels

开源微孔板兼容照明面板的应用

Microplates are commonly used in the modern laboratory environment for a wide variety of tasks both in small-scale laboratory benchtop operations as well ...