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 var en gåva från Kenneth Yamada, National Institutes of Health, National Institute of Dental och Craniofacial Forskning, Bethesda, Maryland, och aktin-mRFP plasmid var en gåva från Michael Davidson, Florida State University, Tallahassee, Florida. Detta arbete stöddes av NSF Karriär utmärkelse # 0747334 och AHA-National SDG # 0835205N till Andreea Trache.

1. Översikt över det integrerade Mikroskop System Mikroskopet system som används för dessa studier har beskrivits i detalj 1. Kortfattat är en inverterad Olympus IX-81 mikroskop med TIRF bilaga (Olympus, Center Valley, PA) i kombination med CSU-22 Yokogawa skanning huvudet (Yokogawa Electric Inc, Japan). En Bioscope SZ atomkraftsmikroskop (Veeco Instruments Inc, Santa Barbara, Kalifornien) är monterad på toppen av den optiska inverterat mikroskop, och hela systemet är placer...

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Cytoskelettet 7-10 och fokus sammanväxningar är 11-16 dynamiska strukturer som monterar, sprida och vänd över som celler migrera eller svarar på mekanisk kraft 17. De har betydande funktioner i regleringen av celltillväxt, överlevnad och genuttryck. Att veta deras specifika distribution och dynamik ger en bättre förståelse för hur celler kommunicerar med matris och mellan sig. Det nya mikroskop systemet är mångsidigt användbar inom ett brett spe...

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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>
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<td>avidin</td>
<td>&nbsp;</td>
<td>Sigma Aldrich</td>
<td>A9275</td>
<td>1mg/mL in DPBS</td>
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<td>Fibronectin</td>
<td>&nbsp;</td>
<td>Sigma Aldrich</td>
<td>F4759</td>
<td>1 mg/mL</td>
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<td>DPBS</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>21600-051</td>
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<td>Amaxa transfection kit for Smooth Muscle Cells</td>
<td>&nbsp;</td>
<td>Lonza</td>
<td>VPI-1004</td>
<td>&nbsp;</td>
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<td>Mattek glass bottom dishes</td>
<td>&nbsp;</td>
<td>MatTek Corporation</td>
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live cell response to mechanical stimulation studied by integrated optical and atomic force microscopy

För att förstå den mekanism genom vilken levande celler känsla mekaniska krafter och hur de reagerar och anpassa sig till sin miljö, en ny teknik kan undersöka cellerna beteende vid sub-cellnivå med hög tidsmässiga och geografiska utvecklades. Således var ett atomkraftsmikroskop (AFM) integrerad med en total inre reflektion fluorescens (TIRF) mikroskopi och snabbt snurrande skiva (FSD) konfokalmikroskopi. Det integrerade systemet som är brett tillämpbar inom ett brett spektrum av molekylära dynamiska studier i någon anhängare levande celler, vilket gör att direkt optisk avbildning av cell svar på mekanisk stimulering i realtid. Betydande ombildning av aktin filament och fokus sammanväxningar visades på grund av lokal mekanisk stimulering på apikala cellytan som inducerade förändringar i cellstrukturen i hela cellkroppen. Dessa innovativa tekniker kommer att ge ny information för att förstå levande cell omstrukturering och dynamik som svar på mekanisk kraft. En detaljerad protokoll och en representant datauppsättning som visar levande cell svar på mekanisk stimulering presenteras.

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

Detta dokument syftar till att instruera läsaren i driften av ett integrerat atomic force-optisk avbildning mikroskop för mekanisk stimulering av levande celler i kultur. En steg-för-steg-protokollet presenteras. En representant datauppsättning som visar levande cell svar på mekanisk stimulering presenteras.

Levande cell Svar på mekanisk stimulering studerats av integrerad optisk och atomkraftsmikroskopi

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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