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

इस काम के हिस्से में दक्षिण टेक्सास प्रौद्योगिकी प्रबंधन (2009.041 POCrr), और अनुसंधान संसाधन के लिए राष्ट्रीय केंद्र (1RR025767 उल) से चिकित्सा विज्ञान के एकीकरण के लिए संस्थान से अनुदान द्वारा वित्त पोषित किया गया था और दंत चिकित्सा और craniofacial रिसर्च के राष्ट्रीय संस्थान (से 5R21DE017294).

1. Functionalized स्लाइड्स की तैयारी <ol><li> एक हटाने योग्य स्लाइड रैक में खुर्दबीन स्लाइड प्लेस, और धो दो बार एक धुंधला 99% इथेनॉल (histological ग्रेड) युक्त जार में डुबो कर. पोछो स्लाइड कागज कागज धूल उत्पन्न नहीं यह सुनिश्चित ...

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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>

हम एक सेल आधारित उच्च घनत्व माइक्रोएरे, CA BChip, Candida albicans के biofilms के nanoliter संस्करणों से मिलकर विकसित किया है. माइक्रोएरे संशोधित ग्लास substrates, जो कोलेजन जेल स्थलों में से एक मजबूत लगाव के लिए अनुमति दी, जबकि एक...

<table border="1"><tbody><tr><td> अभिकर्मक का नाम </td><td> कंपनी </td><td> सूचीपत्र संख्या </td></tr><tr><td> 3 - aminopropyltriethoxysilane (APTES) </td><td> सिग्मा Aldrich </td><td> 440,140 </td></tr><tr><td> Polystyrene - सह - Maleic Anhydride (PS एमए) </td><td> सिग्मा Aldrich </td><td> 426,946 </td></tr><tr><td> ग्लास माइक्रोस्कोपी स्लाइड </td><td> फिशर साइंटिफिक </td><td> 12-549-3 </td></tr><tr><td> मैं चूहा पूँछ कोलेजन टाइप </td><td> बी.डी. बायोसाइंसेज </td><td> 354,236 </td></tr><tr><td> रोबोट Microarrayer </td><td> Omnigrid माइक्रो </td><td> MICROSYS4000/4100A </td></tr><tr><td> माइक्रोएरे स्कैनर </td><td> Genepix निजी 4100A </td><td> GENEPIX4100A </td></tr><tr><td> संकरण कैसेट्स </td><td> ArrayIt निगम </td><td> AHCXD </td></tr><tr><td> FUN1 [2-Chloro-4 (2,3 - dihydro 3 - मिथाइल (benzo -1,3 thiazol-2-YL) - methylidene) 1 फिनाइल quinoliniumiodide] </td><td> Invitrogen कॉर्प </td><td> F-+७०३० </td></tr><tr><td> Fluconazole </td><td> Sicor फार्मास्यूटिकल्स इंक, </td><td> J02AC01 </td></tr><tr><td> Amphotericin बी </td><td> सिग्मा </td><td> A2411 </td></tr><tr><td> RPMI - 1640 </td><td> Mediatech इंक, </td><td> 50-020 - पीसी </td></tr><tr><td> सिरेमिक युक्ति 190 माइक्रोन छिद्र </td><td> Digilab </td><td> 60020441-00 </td></tr><tr><td> GraphPad चश्मे सॉफ्टवेयर </td><td> GraphPad सॉफ्टवेयर इंक, </td><td></td></tr><tr><td> Genepix प्रो v4.1 </td><td> आणविक डिवाइसेज </td><td></td></tr></tbody></table>

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

Candida albicans remains the main etiological agent of candidiasis, which currently represents the fourth most common nosocomial bloodstream infection in US hospitals1. These opportunistic infections pose a growing threat for an increasing number of compromised individuals, and carry unacceptably high mortality rates. This is in part due to the limited arsenal of antifungal drugs, but also to the emergence of resistance against the most commonly used antifungal agents. Further complicating treatment is the fact that a majority of manifestations of candidiasis are associated with the formation of biofilms, and cells within these biofilms show increased levels of resistance to most clinically-used antifungal agents2. Here we describe the development of a high-density microarray that consists of C. albicans nano-biofilms, which we have named CaBChip3. Briefly, a robotic microarrayer is used to print yeast cells of C. albicans onto a solid substrate. During printing, the yeast cells are enclosed in a three dimensional matrix using a volume as low as 50 nL and immobilized on a glass substrate with a suitable coating. After initial printing, the slides are incubated at 37 &deg;C for 24 hours to allow for biofilm development. During this period the spots grow into fully developed "nano-biofilms" that display typical structural and phenotypic characteristics associated with mature C. albicans biofilms (i.e. morphological complexity, three dimensional architecture and drug resistance)4. Overall, the CaBChip is composed of ~750 equivalent and spatially distinct biofilms; with the additional advantage that multiple chips can be printed and processed simultaneously. Cell viability is estimated by measuring the fluorescent intensity of FUN1 metabolic stain using a microarray scanner. This fungal chip is ideally suited for use in true high-throughput screening for antifungal drug discovery. Compared to current standards (i.e. the 96-well microtiter plate model of biofilm formation5), the main advantages of the fungal biofilm chip are automation, miniaturization, savings in amount and cost of reagents and analyses time, as well as the elimination of labor intensive steps. We believe that such chip will significantly speed up the antifungal drug discovery process.

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

हम एक उच्च घनत्व माइक्रोएरे मंच 3 डी नैनो biofilms से मिलकर विकसित किया है<em&gt; सी. सफेद</em&gt; कहा जाता है<em&gt; Ca</emमें&gt; BChip. दवाओं की संवेदनशीलता प्रोफ़ाइल पर परीक्षण किया<em&gt; Ca</em&gt; BChip 96 अच्छी तरह से पारंपरिक थाली मॉडल के लिए तुलनीय है, सुझाव है कि कवक चिप आदर्श रोधी दवाओं का सच उच्च throughput स्क्रीनिंग के लिए अनुकूल है.

उच्च throughput रोधी दवा स्क्रीनिंग के लिए candida albicans के Biofilm चिप (सीए BChip,)

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

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

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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 ...

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बैक्टीरिया के उच्च throughput पहचान चिकित्सा उपकरणों के लिए पॉलिमर से बचाने वाली क्रीम

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

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

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