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">This work was supported by a RO1 from NIH and Merit Reviews from the VA Research Service to C.-M. T., and an individual NRSA to C.W.L.</p>

<p class="jove_title">General design considerations</p>
<p class="jove_content">If the wavelength of photo stimulation is in the visible range, such as for optogenetic experiments, the layout of the system will be considerably simpler than for UV photolysis experiments. One only needs to purchase a dual camera port module that is available from all of the microscope companies. The DMD plane can be positioned at the conjugate image plane of either one of the two camera ports. But in UV photolysis the light originating from the DMD must be brough...

<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">The advantage of DMD based photostimulation approach is most apparent for situations where the target occupies a relatively large area. If the target of interest is very small, such as a few dendritic spines, sequential scanning confocal and 2-photon systems are likely to be the better approach. One significant weakness of the DMD approach is its inefficient use of the available light. The bulk of the available light is necessarily directed to the OFF mirrors and not used.</p>
<p class="jove_content">The DMD based syste...

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

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		<th>Catalogue Number</th>
		<th>Comment</th>
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<tr>
<td>Modern upright fluorescent microscope</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>CCD camera and image acquisition software</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Computer and data acquisition/interface system</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>DLP Discovery Developer Kit </td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>ALP3 USB interface</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>S2 + Optics w/LED</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Dual camera port unit</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</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>
<td>&nbsp;</td>
<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>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>up to 50 kHz</td>
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<td>Beam collimator</td>
<td>&nbsp;</td>
<td>Olympus</td>
<td>DApo20UV340</td>
<td>&nbsp;</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">Digital micromirror devices (DMD) can generate complex patterns in time and space with which to control neuronal excitability. Issues relevant to the design, construction, and operation of DMD systems are discussed. Such a system enabled the demonstration of non-linear integration across distal dendritic branch points.</p>

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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