Name: patterned photostimulation with digital micromirror devices to investigate dendritic integration across branch points
Uploaded: 2011-03-02T17:30:00
Description: Watch this Scientific Journal Video about Patterned Photostimulation with Digital Micromirror Devices to Investigate Dendritic Integration Across Branch Points at JoVE.com

<p class="jove_content"> Questo lavoro è stato supportato da una RO1 dal NIH e recensioni Merito del Servizio di ricerca VA a C.-MT, e un NRSA individuale a CWL </p>

<p class="jove_title"> Considerazioni generali di progettazione </p><p class="jove_content"> Se la lunghezza d&#39;onda di stimolazione foto è nel campo del visibile, come per gli esperimenti optogenetic, il layout del sistema sarà notevolmente più semplice per gli esperimenti di fotolisi UV. Basta acquistare un doppio modulo porta macchina fotografica che è disponibile presso tutte le aziende microscopio. L&#39;aereo DMD può essere posizionato sul piano dell&#39;immagine sia coniugato di una delle due porte della fotocamera. Ma nella foto...

<ol>
	<li>Scanziani, M., Hausser, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiology+in+the+age+of+light.">Electrophysiology in the age of light.</a> <em style='font-style: italic'>Nature</em>. <b>461</b>, 930-939 (2009).</li><li>Tang, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photolysis+of+caged+neurotransmitters:+Theory+and+procedures+for+light+delivery.">Photolysis of caged neurotransmitters: Theory and procedures for light delivery.</a> <em style='font-style: italic'>Curr. Prot. Neurosci</em>. , 6.21.1-6.21.12 (2006).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nature+Technology+Feature,+Cell+imaging:+Light+activated.">Nature Technology Feature, Cell imaging: Light activated.</a> <em style='font-style: italic'>Nature</em>. <b>456</b>, 826-827 (2008).</li><li>Lutz, C., Otis, T. S., DeSars, V., Charpak, S., DeGregorio, D. A., Emiliani, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Holographic+photolysis+of+caged+neurotransmitters.">Holographic photolysis of caged neurotransmitters.</a> <em style='font-style: italic'>Nature Methods</em>. <b>5</b>, 821-827 (2008).</li><li>Hornbeck, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> <em style='font-style: italic'>Digital Light Processing and MEMs: An overview</em>. , .</li></ol>

<p class="jove_content"> Il vantaggio di DMD approccio basato fotostimolazione è più evidente nelle situazioni in cui il target occupa un&#39;area relativamente grande. Se il bersaglio di interesse è molto piccolo, come un paio di spine dendritiche, sistemi di scansione sequenziale confocale e 2-fotoni possono essere l&#39;approccio migliore. Un punto debole significativo dell&#39;approccio DMD è il suo uso inefficiente della luce disponibile. La maggior parte della luce disponibile è necessariamente rivolta al specchi OFF e non utilizzati...

Erratum: Patterned Photostimulation with Digital Micromirror Devices to Investigate Dendritic Integration Across Branch Points

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<td>Modern upright fluorescent microscope</td>
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<td>CCD camera and image acquisition software</td>
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<td>Computer and data acquisition/interface system</td>
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<td>DLP Discovery Developer Kit </td>
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<td>ALP3 USB interface</td>
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<td>S2 + Optics w/LED</td>
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<td>Dual camera port unit</td>
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<td>355nm frequency tripled NdVO4 laser (~1 W)</td>
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<td>DPSS Laser Inc.</td>
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<td>Laser shutter Model LS6</td>
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<td>Uniblitz</td>
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<td>Multimode optical fiber and fiber stretcher Model# 915</td>
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<td>Canadian Instrument and Research, Ltd </td>
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<td>100 um core multimode fiber</td>
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<td>Multimode Fiber launcher </td>
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<td>Oz Optics</td>
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<td>Signal generator</td>
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<td>up to 50 kHz</td>
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<td>Beam collimator</td>
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<td>Olympus</td>
<td>DApo20UV340</td>
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<td>UV relay lens</td>
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<td>Special Optics</td>
<td>#: 54-25-60-355</td>
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patterned photostimulation with digital micromirror devices to investigate dendritic integration across branch points

<p class="jove_content">Light is a versatile and precise means to control neuronal excitability. The recent introduction of light sensitive effectors such as channel-rhodopsin and caged neurotransmitters have led to interests in developing better means to control patterns of light in space and time that are useful for experimental neuroscience. One conventional strategy, employed in confocal and 2-photon microscopy, is to focus light to a diffraction limited spot and then scan that single spot sequentially over the region of interest. This approach becomes problematic if large areas have to be stimulated within a brief time window, a problem more applicable to photostimulation than for imaging. An alternate strategy is to project the complete spatial pattern on the target with the aid of a digital micromirror device (DMD). The DMD approach is appealing because the hardware components are relatively inexpensive and is supported by commercial interests. Because such a system is not available for upright microscopes, we will discuss the critical issues in the construction and operations of such a DMD system. Even though we will be primarily describing the construction of the system for UV photolysis, the modifications for building the much simpler visible light system for optogenetic experiments will also be provided.  The UV photolysis system was used to carryout experiments to study a fundamental question in neuroscience, how are spatially distributed inputs integrated across distal dendritic branch points. The results suggest that integration can be non-linear across branch points and the supralinearity is largely mediated by NMDA receptors.</p>

Watch this Scientific Journal Video about Patterned Photostimulation with Digital Micromirror Devices to Investigate Dendritic Integration Across Branch Points at JoVE.com

Patterned Photostimulation with Digital Micromirror Devices to Investigate Dendritic Integration Across Branch Points

<p class="jove_content">Dispositivi digitali micromirror (DMD) in grado di generare modelli complessi nel tempo e nello spazio con cui controllare l&#39;eccitabilità neuronale. Questioni rilevanti per la progettazione, costruzione e funzionamento dei sistemi di DMD sono discussi. Tale sistema ha permesso la dimostrazione di non-lineare integrazione tra distale punti di ramificazione dendritica.</p>

Fotostimolazione modellato con i dispositivi digitali Micromirror per Indagare integrazione dendritiche Attraverso punti di ramificazione

Light is a versatile and precise means to control neuronal excitability. The recent introduction of light sensitive effectors such as channel-rhodopsin ...

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