Name: live cell response to mechanical stimulation studied by integrated optical and atomic force microscopy
Uploaded: 2010-10-04T17:00:00
Description: Watch this Scientific Journal Video about Live Cell Response to Mechanical Stimulation Studied by Integrated Optical and Atomic Force Microscopy at JoVE.com

Vinculin-GFP-Plasmid war ein Geschenk von Kenneth Yamada, National Institutes of Health, National Institute of Dental und kraniofaziale Research, Bethesda, Maryland, und Aktin-mRFP Plasmid war ein Geschenk von Michael Davidson, Florida State University, Tallahassee, Florida. Diese Arbeit wurde von der NSF Career Award # 0747334 und AHA-National SDG # 0835205N zu Andreea Trache unterstützt.

1. Übersicht über die Integrierte Mikroskop-System Das Mikroskop-System, das für diese Studien verwendet wird, hat in detail 1 beschrieben worden. Kurz gesagt, ist eine umgekehrte Olympus IX-81-Mikroskop mit TIRF Befestigung (Olympus, Center Valley, PA) in Kombination mit CSU-22 Yokogawa Abtastkopf (Yokogawa Electric Inc, Japan). Ein Bioscope SZ Rasterkraftmikroskop (Veeco Instruments Inc, Santa Barbara, CA) ist auf der Oberseite des optischen inverse Mikroskop montiert, und das ...

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Zytoskelett 7-10 und 11-16 fokalen Adhäsionen sind dynamische Strukturen, die, montieren verteilen und umdrehen, wie Zellen wandern oder reagieren auf mechanische Kraft 17. Sie haben wichtige Funktionen bei der Regulation des Zellwachstums, das Überleben und die Genexpression. Das Wissen ihrer spezifischen Verteilung und Dynamik ermöglicht ein besseres Verständnis, wie Zellen mit der Matrix und untereinander zu kommunizieren. Das neue Mikroskop-System ist...

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		<th>Material Name</th>
		<th>Type</th>
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		<th>Catalogue Number</th>
		<th>Comment</th>
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<td>AFM tip</td>
<td>&nbsp;</td>
<td>Novascan Technologies</td>
<td>&nbsp;</td>
<td>2 micron Borosilicate bead, 0.01N/m, Biotin Surface</td>
</tr>
<tr>
<td>avidin</td>
<td>&nbsp;</td>
<td>Sigma Aldrich</td>
<td>A9275</td>
<td>1mg/mL in DPBS</td>
</tr>
<tr>
<td>Fibronectin</td>
<td>&nbsp;</td>
<td>Sigma Aldrich</td>
<td>F4759</td>
<td>1 mg/mL</td>
</tr>
<tr>
<td>DPBS</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>21600-051</td>
<td>&nbsp;</td>
<tr>
<td>Amaxa transfection kit for Smooth Muscle Cells</td>
<td>&nbsp;</td>
<td>Lonza</td>
<td>VPI-1004</td>
<td>&nbsp;</td>
<tr>
<td>Mattek glass bottom dishes</td>
<td>&nbsp;</td>
<td>MatTek Corporation</td>
<td>P60G-1.5-30-F</td>
<td>Sterile 60mm Falcon dishes with glass bottom coverslip no. 1.5</td>
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live cell response to mechanical stimulation studied by integrated optical and atomic force microscopy

Um zu verstehen, den Mechanismus, mit dem sich lebende Zellen Sinn mechanische Kräfte, und wie sie zu reagieren und an ihre Umgebung anpassen, eine neue Technologie in der Lage, Zellen Verhalten bei sub-zellulärer Ebene zu untersuchen mit hoher räumlicher und zeitlicher Auflösung entwickelt wurde. So wurde ein Rasterkraftmikroskop (AFM) mit Total Internal Reflection Fluorescence (TIRF) Mikroskopie und schnell drehenden Scheibe (FSD) konfokale Mikroskopie integriert. Das integrierte System ist breit anwendbar für eine Vielzahl von molekularen dynamischen Studien in anhaftendem lebenden Zellen, die eine direkte optische Bildgebung von Zell-Reaktionen auf mechanische Reize in Echtzeit. Signifikante Umlagerung des Aktin-Filamente und fokalen Adhäsionen war aufgrund der örtlichen mechanische Stimulation an der apikalen Zelloberfläche, dass Veränderungen in der Zellstruktur im gesamten Zellkörper induzierte gezeigt. Diese innovative Technik wird neue Informationen zum Verständnis lebender Zellen Umstrukturierung und Dynamik in Reaktion auf die mechanische Kraft. Ein detailliertes Protokoll sowie ein Vertreter Datensatz, zeigen Live-Zell-Antwort auf mechanische Reize präsentiert werden.

Watch this Scientific Journal Video about Live Cell Response to Mechanical Stimulation Studied by Integrated Optical and Atomic Force Microscopy at JoVE.com

Live Cell Response to Mechanical Stimulation Studied by Integrated Optical and Atomic Force Microscopy

Dieses Papier soll den Leser in den Betrieb eines integrierten atomic force-optische Bildgebung Mikroskop für die mechanische Stimulation von lebenden Zellen in Kultur zu unterrichten. Eine Schritt-für-Schritt-Protokoll vorgestellt. Eine repräsentative Datenmenge, die Live-Zell-Antwort auf mechanische Stimulation zeigt vorgestellt.

Live Cell Reaktion auf mechanische Stimulation von Integrated Optical und Atomic Force Microscopy Studium

To understand the mechanism by which living cells sense mechanical forces, and how they respond and adapt to their environment, a new technology able to ...

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