Name: measurement of bioelectric current with a vibrating probe
Uploaded: 2011-01-04T17:00:00
Description: Watch this Scientific Journal Video about Measurement of Bioelectric Current with a Vibrating Probe at JoVE.com

Siamo grati al professor Richard Borgens, Centro per la Ricerca Paralisi, Purdue University, per un aiuto per il montaggio della sonda sistema vibrante. Questo studio è stato sostenuto da NIH 1R01EY019101 NEI concedere a MZ e BR, e in parte da finanziamenti del California Institute of Regenerative Medicine RB1-01417, NSF MCB-0951199, e da una sovvenzione illimitata di ricerca per prevenire la cecità, UC Davis Oftalmologia.

1. Sonda Produzione Sonde vuote sono acquistati da Strumenti di precisione (World Elgiloy / acciaio rivestita parylene microelettrodi) (vedi &#39;Tabella di reagenti e attrezzature specifiche&#39; di seguito). La sonda viene tagliata 25-30 mm dietro la punta e di circa 5 mm di isolamento parylene alla fine taglio raschiato via con un bisturi (# 11 della lama) per garantire una buona connessione. La sonda è montata in un oro R30 connettore utilizzando elettricamente conduttivo d&#39;argento ca...

<ol>
	<li>Reid, B., Nuccitelli, R., Zhao, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+measurement+of+bioelectric+currents+with+a+vibrating+probe.">Non-invasive measurement of bioelectric currents with a vibrating probe.</a> Nat. Protoc. 2, 661-9282 (2007).</li><li>Reid, B., Song, B., McCaig, C. D., Zhao, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wound+healing+in+rat+cornea:+the+role+of+electric+currents.">Wound healing in rat cornea: the role of electric currents.</a> FASEB J. 19, 379-386 (2005).</li><li>Lois, N., Reid, B., Song, B., Zhao, M., Forrester, J. V., McCaig, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electric+currents+and+lens+regeneration+in+the+rat.">Electric currents and lens regeneration in the rat.</a> Exp. Eye Res. 90, 316-323 (2010).</li><li>Wang, E., Reid, B., Lois, N., Forrester, J. V., McCaig, C. D., Zhao, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+inhibition+of+lens+epithelial+cell+proliferation:+an+additional+factor+in+secondary+cataract.">Electrical inhibition of lens epithelial cell proliferation: an additional factor in secondary cataract.</a> FASEB J. 19, 842-844 (2005).</li><li>Guo, A., Song, B., Reid, B., Gu, Y., Forrester, J. V., Jahoda, C., Zhao, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+physiological+electric+fields+on+migration+of+human+dermal+fibroblasts.">Effects of physiological electric fields on migration of human dermal fibroblasts.</a> J. Invest. Derm. , (2010).</li><li>Reid, B., Song, B., Zhao, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electric+currents+in+Xenopus+tadpole+tail+regeneration.">Electric currents in Xenopus tadpole tail regeneration.</a> Dev. Biol. 335, 198-207 (2009).</li><li>Zhao, M., Song, B., Pu, J., Wada, T., Reid, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+signals+control+wound+healing+through+phosphatidylinositol-3-OH+kinase-&#947;.">Electrical signals control wound healing through phosphatidylinositol-3-OH kinase-&#947;.</a> 442, 457-460 (2006).</li><li>Reid, B., EO, G. r. a. u. e. -. H. e. r. n. a. n. d. e. z., Mannis, M. J., Zhao, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulating+endogenous+electric+currents+in+human+corneal+wounds+-+a+novel+approach+of+bioelectric+stimulation+without+electrodes.">Modulating endogenous electric currents in human corneal wounds - a novel approach of bioelectric stimulation without electrodes.</a> Cornea. , (2010).</li><li>Nuccitelli, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+ultrasensitive+vibrating+probe+for+measuring+steady+extracellular+currents.">An ultrasensitive vibrating probe for measuring steady extracellular currents.</a> J. Cell Biol. 63, 614-628 (1974).</li><li>Hotary, K. B., Nuccitelli, R., Robinson, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+computerized+2-dimensional+vibrating+probe+for+mapping+extracellular+current+patterns.">A computerized 2-dimensional vibrating probe for mapping extracellular current patterns.</a> J. Neurosci. Meth. 43, 55-67 (1992).</li><li>Nuccitelli, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endogenous+ion+currents+and+DC+electric+fields+in+multicellular+animal+tissues.">Endogenous ion currents and DC electric fields in multicellular animal tissues.</a> Bioelectromagnetics Supplement. 1, 147-157 (1992).</li><li>Levin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioelectric+mechanisms+in+regeneration:+Unique+aspects+and+future+perspectives.">Bioelectric mechanisms in regeneration: Unique aspects and future perspectives.</a> Seminars in Cell Dev. Biol. 20, 543-556 (2009).</li><li>Zhao, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electric+fields+in+wound+healing+-+An+overriding+signal+that+directs+cell+migration.">Electric fields in wound healing - An overriding signal that directs cell migration.</a> Seminars in Cell Dev. Biol. 20, 674-682 (2009).</li></ol>

Descriviamo un basso costo, di base, ma molto sensibile sistema sonda vibrante per la misurazione non invasiva corrente elettrica in una varietà di sistemi biologici. Le modifiche possibili <ol><li> Se platino / iridio elettrodi (Strumenti di precisione mondiale; cat # PTM23B20) sono utilizzati al posto di acciaio inox, poi la fase di placcatura in oro può essere eliminato. </li></ol> Applicazioni Abbiamo usato la sonda vibrante per misura...

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		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Eligoy-Stainless Electrode</td>
<td>&nbsp;</td>
<td>World Precision Instruments</td>
<td>SSM33A70</td>
<td>76 mm, 7 M&Omega;, 1-2&mu;m tip</td>
</tr>
<tr>
<td>Gold R30 connector</td>
<td>&nbsp;</td>
<td>www.vectorelect.com</td>
<td>R30</td>
<td>Re-usable</td>
</tr>
<tr>
<td>Silver-loaded epoxy</td>
<td>&nbsp;</td>
<td>3M</td>
<td>SL65</td>
<td>Mix 1-part Resin with 1-part Hardener</td>
</tr>
<tr>
<td>Dissecting microscope</td>
<td>&nbsp;</td>
<td>Olympus</td>
<td>SZ40</td>
<td>Magnification x6 to x40</td>
</tr>
<tr>
<td>Potassium dicyanoaurate (KAu(CN)2)</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>379867</td>
<td>CAUTION: Toxic</td>
</tr>
<tr>
<td>Chloroplatinic acid hydrate (H2PtCl6 x 6H2O)</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>520896</td>
<td>CAUTION: Toxic</td>
</tr>
<tr>
<td>Lead(II) acetate trihydrate (Pb(CH3CO2)2 x 3H2O)</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>185191</td>
<td>CAUTION: Toxic</td>
</tr>
<tr>
<td>Nano-Amp power source</td>
<td>&nbsp;</td>
<td>Made in-house</td>
<td>-</td>
<td>Powered by six 1.5 V (AAA) batteries</td>
</tr>
<tr>
<td>3-dimensional micro-positioner</td>
<td>&nbsp;</td>
<td>Line Tool Co.</td>
<td>Model H</td>
<td>&nbsp;</td>
<tr>
<td>Lock-in amplifier</td>
<td>&nbsp;</td>
<td>Stanford Research Systems</td>
<td>SR530</td>
<td>&nbsp;</td>
<tr>
<td>Digital I/O interface</td>
<td>&nbsp;</td>
<td>National instruments</td>
<td>PCI-6220</td>
<td>&nbsp;</td>
<tr>
<td>Shielded Connector Block with BNC connections</td>
<td>&nbsp;</td>
<td>National instruments</td>
<td>BNC-2110</td>
<td>&nbsp;</td>
<tr>
<td>Strathclyde Electrophysiology Software</td>
<td>&nbsp;</td>
<td>University of Strathclyde Institute of Pharmacy and Biomedical Sciences, UK</td>
<td>WinWCP V4.1.5</td>
<td>Free download from: <a href="http://spider.science.strath.ac.uk/sipbs/software_ses.htm">http://spider.science.strath.ac.uk/sipbs/software_ses.htm</a></td>
</tr>
<tr>
<td>Calibration Chamber</td>
<td>&nbsp;</td>
<td>Made in-house</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Constant Current Calibrator</td>
<td>&nbsp;</td>
<td>Vibrating Probe Company</td>
<td>&nbsp;</td>
<td>Powered by one 9 V (PP3) battery</td>
</tr>		</tbody>
		</table>
		</div>

measurement of bioelectric current with a vibrating probe

Campi elettrici, generati dal trasporto attivo di ioni, sono presenti in molti sistemi biologici e spesso svolgono funzioni importanti in tessuti e organi. Ad esempio, essi svolgono un ruolo importante nel dirigere la migrazione delle cellule durante la guarigione delle ferite. Qui si descrive la fabbricazione e l&#39;uso di sonde a vibrazione ultrasensibile per la misura extracellulare correnti elettriche. Il sensore è un isolato, filo metallico affilato con una piccola punta di platino-nero (30-35 micron), in grado di rilevare le correnti ioniche nei ìA / cm 2 gamma di soluzione fisiologica. La sonda viene fatta vibrare a circa 200 Hz da un piezoelettrico bender. In presenza di una corrente ionica, la sonda rileva una differenza di tensione tra gli estremi del suo movimento. Un lock-in filtri amplificatore i rumori estranei, bloccando alla frequenza della sonda di vibrazione. I dati vengono registrati su computer. La sonda è calibrata all&#39;inizio e alla fine degli esperimenti in soluzione fisiologica del caso, con una camera che si applica una corrente di esattamente 1,5 ìA / cm 2. Noi descriviamo come fare le sonde, impostare il sistema e calibrare. Abbiamo anche dimostrare la tecnica di misurazione della cornea, e mostrano alcuni risultati rappresentante di diversi esemplari (cornea, cute, cervello).

Watch this Scientific Journal Video about Measurement of Bioelectric Current with a Vibrating Probe at JoVE.com

Measurement of Bioelectric Current with a Vibrating Probe

L&#39;, taratura fabbricazione e l&#39;uso di sonde a vibrazione non invasivo per misurare la corrente bioelettrica in vari sistemi biologici è descritto.

Misura della corrente bioelettrica con una sonda vibrante

Electric fields, generated by active transport of ions, are present in many biological systems and often serve important functions in tissues and organs. ...

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