Name: fabrication and use of microenvironment microarrays (mearrays)
Uploaded: 2012-10-11T12:00:00
Description: Watch this Scientific Journal Video about Fabrication and Use of MicroEnvironment microArrays MEArrays at JoVE.com

एमएल एनआईए (R00AG033176 और R01AG040081) और प्रयोगशाला के निर्देश के अनुसंधान और विकास, अमेरिका डे AC02 - 05CH11231 # ऊर्जा अनुबंध की विभाग द्वारा समर्थित है.

1. मुद्रण Substrata तैयारी polydimethylsiloxane (PDMS) लेपित या polyacrylamide (पीए) लेपित स्लाइड्स का उपयोग करने का निर्णय प्रयोगात्मक डिजाइन के महत्वपूर्ण मानकों पर निर्भर करता है. दोनों पॉलिमर के लोचदार मापांक PDMS का अनुपा?...

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	<li>Bissell, M. J., Labarge, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Context,+tissue+plasticity,+and+cancer:+are+tumor+stem+cells+also+regulated+by+the+microenvironment.">Context, tissue plasticity, and cancer: are tumor stem cells also regulated by the microenvironment.</a> Cancer Cell. 7, 17-23 (2005).</li><li>Engler, A. J., Sen, S., Sweeney, H. L., Discher, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Matrix+elasticity+directs+stem+cell+lineage+specification.">Matrix elasticity directs stem cell lineage specification.</a> Cell. 126, 677-689 (2006).</li><li>McBeath, R., Pirone, D. M., Nelson, C. M., Bhadriraju, K., Chen, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+shape,+cytoskeletal+tension,+and+RhoA+regulate+stem+cell+lineage+commitment.">Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment.</a> Dev. Cell. 6, 483-495 (2004).</li><li>LaBarge, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+mammary+progenitor+cell+fate+decsions+are+products+of+interactions+with+combinatorial+microenvironments.">Human mammary progenitor cell fate decsions are products of interactions with combinatorial microenvironments.</a> Integrative Biology. 1, 70-79 (2009).</li><li>Kim, H. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterning+Methods+for+Polymers+in+Cell+and+Tissue+Engineering.">Patterning Methods for Polymers in Cell and Tissue Engineering.</a> Annals of biomedical engineering. , (2012).</li><li>Boudou, T., Ohayon, J., Picart, C., Pettigrew, R. I., Tracqui, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonlinear+elastic+properties+of+polyacrylamide+gels:+implications+for+quantification+of+cellular+forces.">Nonlinear elastic properties of polyacrylamide gels: implications for quantification of cellular forces.</a> Biorheology. 46, 191-205 (2009).</li><li>Tse, J. R., Engler, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+hydrogel+substrates+with+tunable+mechanical+properties.">Preparation of hydrogel substrates with tunable mechanical properties.</a> Current protocols in cell biology. Chapter 10, Unit 10 (2010).</li><li>Flaim, C. J., Chien, S., Bhatia, S. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+extracellular+matrix+microarray+for+probing+cellular+differentiation.">An extracellular matrix microarray for probing cellular differentiation.</a> Nat. Methods. 2, 119-125 (2005).</li><li>Soen, Y., Mori, A., Palmer, T. D., Brown, P. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exploring+the+regulation+of+human+neural+precursor+cell+differentiation+using+arrays+of+signaling+microenvironments.">Exploring the regulation of human neural precursor cell differentiation using arrays of signaling microenvironments.</a> Mol. Syst. Biol. 2, 37 (2006).</li></ol>

MEArray यहाँ प्रस्तुत विधि सेल और मिश्रित microenvironment बातचीत 4 के कार्यात्मक विश्लेषण के लिए सक्षम बनाता है. इस MEArray विश्लेषण बुनियादी micropatterning प्रौद्योगिकियों, कोशिका जीव विज्ञान, और मुद्रण माइक्रोएरे रोबोट औ?...

<table border="1"><tbody><tr><td> अभिकर्मक के नाम </td><td> कंपनी </td><td> कैटलॉग संख्या </td><td> टिप्पणियां (वैकल्पिक) </td></tr><tr><td> ग्लास 25 मिमी x 75 मिमी स्लाइड </td><td> VWR </td><td> 48311-600 </td><td></td></tr><tr><td> ग्लास (नंबर 1) 24 मिमी x 50 मिमी coverslips </td><td> VWR </td><td> 48393-241 </td><td></td></tr><tr><td> धुंधला डिश (या Coplan जार) </td><td> VWR </td><td> 25461-003 </td><td></td></tr><tr><td> पेट्री डिश (15 सेमी) </td><td> BD फाल्कन </td><td> 351058 </td><td></td></tr><tr><td> NaOH (1.0N) </td><td> सिग्मा Aldrich </td><td> S2567 </td><td></td></tr><tr><td> वानर (&gt; 98% (3-Aminopropyl) triethoxysilane) </td><td> सिग्मा Aldrich </td><td> A3648 </td><td></td></tr><tr><td> Glutaraldehyde </td><td> सिग्मा Aldrich </td><td> G7651 </td><td> पानी में 50% </td></tr><tr><td> ए पी एस (&gt; 98% अमोनियम Persulfate) </td><td> एसigma-Aldrich </td><td> A3678 </td><td> DDH 2 हे के साथ 10% काम समाधान तैयार </td></tr><tr><td> TEMED (एन, एन, एन &#39;, n&#39; Tetramethylethylenediamine) </td><td> सिग्मा Aldrich </td><td> T9281 </td><td></td></tr><tr><td> Acrylamide (40%) </td><td> सिग्मा Aldrich </td><td> A4058 </td><td></td></tr><tr><td> बीआईएस Acrylamide (2% w / v) </td><td> फिशर BioReagents </td><td> BP1404-250 </td><td></td></tr><tr><td> 0.45 सुक्ष्ममापी सिरिंज फिल्टर नायलॉन 4 मिमी </td><td> Nalgene </td><td> 176-0045 </td><td></td></tr><tr><td> FITC </td><td> सिग्मा Aldrich </td><td> F4274 </td><td></td></tr><tr><td> PDMS (polydimethylsiloxane) </td><td> डॉव Corning </td><td> 3097358-1004 </td><td> Sylgard 184 Ellsworth चिपकने के माध्यम से Elastomer किट </td></tr><tr><td> 2 चैम्बर स्लाइड्स </td><td> NUNC </td><td> 177380 </td><td></td></tr><tr><td> F108 Pluronic </td><td> BASF </td><td> 30089186 </td><td></td></tr><tr><td> Aquसंज्ञावाचक शब्दों में लगनेवाला प्रत्यय, यह प्राय: प्राचीन शब्दों में लगता है सीलेंट </td><td> डॉव Corning </td><td> 00688 डीएपी </td><td></td></tr><tr><td> Fluormount-G </td><td> दक्षिणी बायोटेक </td><td> 0100-01 </td><td></td></tr><tr><td> डिस्पोजेबल प्लास्टिक के कप </td><td></td><td></td><td></td></tr><tr><td> जीभ depressors </td><td></td><td></td><td></td></tr><tr><td> Nitrile दस्ताने </td><td></td><td></td><td></td></tr><tr><td> प्लास्टिक खुर्दबीन स्लाइड बक्से </td><td></td><td></td><td></td></tr><tr><td> स्पिन coater </td><td> WS-400B-6NPP/LITE </td><td> Laurell टेक्नोलॉजीज निगम </td><td></td></tr><tr><td> ओवन </td><td></td><td></td><td></td></tr><tr><td> डिजिटल hotplate </td><td></td><td></td><td></td></tr><tr><td> 384 अच्छी तरह प्लेटें </td><td></td><td></td><td> एक माइक्रोएरे रोबोट के लिए उपयुक्त ब्रांड </td></tr><tr><td> मुद्रण माइक्रोएरे रोबोट </td><td></td><td></td><td></td></tr><tr><td> उल्टे चरण और प्रतिदीप्ति सूक्ष्मदर्शी </td><td></td><td></td><td></td></tr><tr><td> Axon माइक्रोएरे स्कैनर </td><td> आणविक उपकरणों </td><td></td><td> एकाधिक विन्यास मौजूद </td></tr></tbody></table>

fabrication and use of microenvironment microarrays (mearrays)

The interactions between cells and their surrounding microenvironment have functional consequences for cellular behaviour. On the single cell level, distinct microenvironments can impose differentiation, migration, and proliferation phenotypes, and on the tissue level the microenvironment processes as complex as morphogenesis and tumorigenesis1. Not only do the cell and molecular contents of microenvironments impact the cells within, but so do the elasticity2 and geometry3 of the tissue. Defined as the sum total of cell-cell, -ECM, and -soluble factor interactions, in addition to physical characteristics, the microenvironment is complex. The phenotypes of cells within a tissue are partially due to their genomic content and partially due to the combinatorial interactions with the microenviroment. A major challenge is to link specific combinations of microenvironmental components with distinctive behaviours.
Here, we present the microenvironment microarray (MEArray) platform for cell-based functional screening of interactions with combinatorial microenvironments4. The method allows for simultaneous control of the molecular composition and the elastic modulus, and combines the use of widely available microarray and micropatterning technologies. MEArray screens require as few as 10,000 cells per array, which facilitates functional studies of rare cell types such as adult progenitor cells. A limitation of the technology is that entire tissue microenvironments cannot be completely recapitulated on MEArrays. However, comparison of responses in the same cell type to numerous related microenvironments, for instance pairwise combinations of ECM proteins that characterize a given tissue, will provide insights into how microenvironmental components elicit tissue-specific functional phenotypes.
MEArrays can be printed using a wide variety of recombinant growth factors, cytokines, and purified ECM proteins, and combinations thereof. The platform is limited only by the availability of specific reagents. MEArrays are amenable to time-lapsed analysis, but most often are used for end point analyses of cellular functions that are measureable with fluorescent probes. For instance, DNA synthesis, apoptosis, acquisition of differentiated states, or production of specific gene products are commonly measured. Briefly, the basic flow of an MEArray experiment is to prepare slides coated with printing substrata and to prepare the master plate of proteins that are to be printed. Then the arrays are printed with a microarray robot, cells are allowed to attach, grow in culture, and then are chemically fixed upon reaching the experimental endpoint. Fluorescent or colorimetric assays, imaged with traditional microscopes or microarray scanners, are used to reveal relevant molecular and cellular phenotypes (Figure 1).

Watch this Scientific Journal Video about Fabrication and Use of MicroEnvironment microArrays MEArrays at JoVE.com

Fabrication and Use of MicroEnvironment microArrays MEArrays

एक मिश्रित सेलुलर कार्यों पर microenvironments की आणविक संरचना के प्रभावों में अंतर्दृष्टि पाने के लिए कार्यात्मक स्क्रीनिंग पद्धति में वर्णित है. विधि मौजूदा माइक्रोएरे आधारित प्रौद्योगिकी का लाभ लेता है परिभाषित मिश्रित microenvironments के arrays कि कोशिका आसंजन और कार्यात्मक विश्लेषण का समर्थन उत्पन्न.

निर्माण और microenvironment प्रोटीन के उपयोग (MEArrays)

The interactions between cells and their surrounding microenvironment have functional consequences for cellular behaviour. On the single cell level, ...

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Biology

Fabrication and Use of MicroEnvironment microArrays (MEArrays)

Bacterial Gene Expression Analysis Using Microarrays

बैक्टीरियल जीन एक्सप्रेशन प्रोटीन का उपयोग विश्लेषण

PDMS Device Fabrication and Surface Modification

PDMS डिवाइस निर्माण और भूतल संशोधन

Fabrication of the Thermoplastic Microfluidic Channels

थर्माप्लास्टिक microfluidic चैनलों के निर्माण कार्य

In our lab, we have successfully isolated nucleic acids directly from microliter and submicroliter volumes of human blood, urine and stool using ...

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Carbon fiber electrodes are crucial for the detection of catecholamine release from vesicles in single cells for amperometry measurements.  Here, we ...

Fabrication of Myogenic Engineered Tissue Constructs

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Despite the fact that electronic pacemakers are life-saving medical devices, their long-term performance in pediatric patients can be problematic owing to ...

Genome-wide Analysis of Aminoacylation (Charging) Levels of tRNA Using Microarrays

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जीनोम चौड़ा Aminoacylation tRNA (चार्ज) स्तर प्रोटीन का उपयोग का विश्लेषण

tRNA aminoacylation, or charging, levels can rapidly change within a cell in response to the environment[1]. Changes in tRNA charging levels in both ...

DNA Microarrays: Sample Quality Control, Array Hybridization and Scanning

नमूना गुणवत्ता नियंत्रण, ऐरे संकरण और स्कैन: डीएनए

Microarray expression profiling of the nervous system provides a powerful approach to identifying gene activities in different stages of development, ...

Production of Tissue Microarrays, Immunohistochemistry Staining and Digitalization Within the Human Protein Atlas

ऊतक, immunohistochemistry धुंधला और डिजिटलीकरण की मानव प्रोटीन एटलस के भीतर उत्पादन

The  tissue microarray (TMA) technology provides the means for high-throughput  analysis of multiple tissues and cells. The technique is used within the ...

Glycan Profiling of Plant Cell Wall Polymers using Microarrays

संयंत्र कोशिका दीवार पॉलिमर माइक्रोएरे का उपयोग कर के glycan रूपरेखा

Plant cell walls are complex matrixes of heterogeneous glycans which play an important role in the physiology and development of plants and provide the ...

Design and Use of Multiplexed Chemostat Arrays

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Chemostats are continuous culture systems in which cells are grown in a tightly controlled, chemically constant environment where culture density is ...