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

Vi är tacksamma till professor Richard Borgens, Centrum för Förlamning forskning, Purdue University, för hjälp med montering av vibrerande sond systemet. Denna studie stöddes av NEI bevilja NIH 1R01EY019101 till MZ och BR, dels genom anslag från California Institute of regenerativ medicin RB1-01.417, NSF MCB-0.951.199 och med ett obegränsat Grant från Forskning kring att förebygga blindhet, UC Davis oftalmologi.

1. Probe Tillverkning Tomma givare köps från instrument World Precision (elgiloy / rostfritt Parylen-coated microelectrodes) (se &quot;Tabell över specifika reagenser och utrustning&quot; nedan). Sonden är skuren 25-30 mm bakom spetsen och ca 5 mm Parylen isolering på den avskurna ändan skrapas bort med en skalpell (# 11 blad) för att säkerställa en god anslutning. Sonden är monterad i en guld R30 kontakt med elektriskt ledande silver-laddad epoxi (t.ex. Rite-Lok SL65) [* se kommenta...

<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>

Vi beskriver en låg kostnad, grundläggande, men mycket känslig vibrerande sond system för att mäta icke-invasivt elektrisk ström i olika biologiska system. Eventuella ändringar <ol><li> Om platina / iridium elektroder (World precisionsinstrument, katt # PTM23B20) används i stället för rostfritt stål, då förgyllning skede kan elimineras. </li></ol> Applikationer Vi har använt den vibrerande sond för att mäta elektrisk ström i...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
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<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>
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measurement of bioelectric current with a vibrating probe

Elektriska fält, som genereras av aktiv transport av joner, finns i många biologiska system och tjänar ofta viktiga funktioner i vävnader och organ. Till exempel spelar de en viktig roll i att leda cell migration vid sårläkning. Här beskriver vi tillverkning och användning av ultrakänslig vibrerande givare för att mäta extracellulära elektriska strömmar. Sonden är en isolerad, vass metalltråd med en liten platina-svart spets (30-35 mikrometer), som kan upptäcka jonströmmar i μA / cm 2 intervall i fysiologisk koksaltlösning. Sonden är vibrerat på ca 200 Hz med en piezoelektrisk Bender. I närvaro av en jonisk ström, upptäcker sonden en spänning skillnad mellan ytterligheterna av dess rörelse. Ett lås förstärkare filtrerar bort brus genom att låsa på sondens frekvensen av vibrationer. Data spelas in på datorn. Sonden är kalibrerad i början och slutet av experiment i lämpliga saltlösning, med hjälp av en kammare som gäller en ström på exakt 1,5 μA / cm 2. Vi beskriver hur man gör sonder, ställa in systemet och kalibrera. Vi visar också tekniken att hornhinnan mätning, och visar några representativa resultat från olika prover (hornhinna, hud, hjärna).

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

Tillverkning, kalibrering och användning av icke-invasiv vibrerande prober för mätning av bioelektriska aktuella i olika biologiska system beskrivs.

Mätning av bioelektriska Aktuell med en vibrerande Probe

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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