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

Plasmidique vinculine-GFP a été un cadeau de Kenneth Yamada, National Institutes of Health, Institut national de recherche dentaire et craniofaciale, Bethesda, Maryland, et l&#39;actine-MRFP plasmide a été un cadeau de Michael Davidson, Florida State University, Tallahassee, en Floride. Ce travail a été soutenu par la NSF Career Award # 0747334 et AHA-National SDG # 0835205N d&#39;Andreea Trache.

1. Présentation du système de microscope intégré Le système de microscope qui est utilisé pour ces études a été décrite en détail 1. En bref, une inversé Olympus IX-81 microscope TIRF attachement (Olympus, Center Valley, Pennsylvanie) est combiné avec la CSU-22 Yokogawa numérisation tête (Yokogawa Electric Inc, Japon). Un microscope à force atomique Bioscope SZ (Veeco Instruments Inc, Santa Barbara, Californie) est monté sur le dessus du microscope optique inversé,...

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Adhérences Cytosquelette 7-10 et 11-16 focale sont des structures dynamiques qui s&#39;assemblent, se dispersent et retournent comme des cellules migrent ou répondre à une force mécanique 17. Ils ont des fonctions importantes dans la régulation de la croissance cellulaire, la survie et l&#39;expression des gènes. Connaissant leur distribution spécifique et de la dynamique permet de mieux comprendre comment les cellules communiquent avec la matrice et entre eux. <p class="jove_...

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		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
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<tr>
<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

Pour comprendre le mécanisme par lequel les cellules vivantes sens des forces mécaniques, et comment ils réagissent et s&#39;adaptent à leur environnement, une nouvelle technologie capable de d&#39;étudier le comportement des cellules au niveau sub-cellulaire avec une résolution spatiale et temporelle élevée a été développé. Ainsi, un microscope à force atomique (AFM) a été intégré à la réflexion interne totale de fluorescence (FRBR) microscopie et rotation rapide du disque (DSE) microscopie confocale. Le système intégré est largement applicable dans un large éventail d&#39;études moléculaires dynamiques dans toute cellules adhérentes en direct, permettant d&#39;imagerie optique directe des réponses des cellules à la stimulation mécanique en temps réel. Réarrangement significatif des filaments d&#39;actine et adhésions focales a été montré en raison de la stimulation mécanique locale à la surface cellulaire apicale qui induit des changements dans la structure cellulaire dans le corps cellulaire. Ces techniques innovantes fournira de nouvelles informations pour comprendre la restructuration de cellules vivantes et dynamiques en réponse à une force mécanique. Un protocole détaillé et un ensemble de données représentatives que la réponse cellulaire spectacle à la stimulation mécanique sont présentées.

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

Ce document vise à instruire le lecteur dans le fonctionnement d&#39;un système intégré à force atomique, microscope optique d&#39;imagerie pour la stimulation mécanique des cellules vivantes en culture. Un protocole étape par étape est présentée. Un ensemble représentatif de données qui montre la réponse de cellules vivantes à la stimulation mécanique est présenté.

Réponse Live Cell à la stimulation mécanique étudiée par microscopie à force atomique et optique intégré

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