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<article dtd-version="3.0" article-type="Corporate Submission">
  <front>
    <journal-meta>
      <journal-id journal-id-type="publisher">jove</journal-id>
      <journal-id journal-id-type="email">editorial-office@jove.com</journal-id>
      <journal-title-group>
        <journal-title>Journal of Visualized Experiments</journal-title>
      </journal-title-group>
      <issn pub-type="ppub">1940-087X</issn>
      <issn pub-type="epub"></issn>
    </journal-meta>
    <article-meta>
      <article-id pub-id-type="publisher-id">JoVE58421</article-id>
      <article-id pub-id-type="manuscript">JoVE58421</article-id>
      <article-categories>
        <subj-group subj-group-type="Article Type">
          <subject>Corporate Submission</subject>
        </subj-group>
        <subj-group subj-group-type="Classification">
          <subject>Technology</subject>
          <subject>Miniaturization</subject>
          <subject>Microtechnology</subject>
          <subject>Microfluidics</subject>
          <subject>Culture Techniques</subject>
          <subject>Cell Culture Techniques</subject>
          <subject>biology (general)</subject>
          <subject>life sciences</subject>
          <subject>animal models</subject>
          <subject>bioengineering (general)</subject>
          <subject>Anatomy</subject>
          <subject>Cells</subject>
          <subject>Nervous System</subject>
          <subject>Analytical, Diagnostic and Therapeutic Techniques and Equipment</subject>
          <subject>Investigative Techniques</subject>
          <subject>Technology, Industry, Agriculture</subject>
          <subject>Technology, Industry, and Agriculture</subject>
          <subject>Life sciences</subject>
          <subject>Life Sciences (General)</subject>
        </subj-group>
      </article-categories>
      <title-group>
        <article-title>Use of pre-assembled plastic microfluidic chips for compartmentalizing primary murine neurons</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author">
          <role content-type="1" />
          <name>
            <surname>Nagendran</surname>
            <given-names>Tharkika</given-names>
          </name>
          <email>tharkika@gmail.com</email>
          <xref ref-type="aff" rid="aff1" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="2" />
          <name>
            <surname>Poole</surname>
            <given-names>Valerie</given-names>
          </name>
          <email>vpoole@xona.us</email>
          <xref ref-type="aff" rid="aff2" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="3" />
          <name>
            <surname>Harris</surname>
            <given-names>Joseph</given-names>
          </name>
          <email>jharris@xona.us</email>
          <xref ref-type="aff" rid="aff3" />
        </contrib>
        <contrib contrib-type="author" corresp="yes">
          <role content-type="4" />
          <name>
            <surname>Taylor</surname>
            <given-names>Anne  </given-names>
            <prefix>Dr.</prefix>
          </name>
          <email>ataylor@xona.us</email>
          <xref ref-type="aff" rid="aff4" />
          <xref ref-type="aff" rid="orcid7" />
        </contrib>
        <contrib contrib-type="editor">
          <name>
            <surname>Navani</surname>
            <given-names>Dipesh</given-names>
          </name>
          <degrees>Ph.D.</degrees>
          <email>dipesh.n@jove.com</email>
          <xref ref-type="aff" rid="aff5" />
        </contrib>
        <aff id="aff1">
          <institution>University of North Carolina at Chapel Hill</institution>
          <addr-line content-type="addrline1">UNC Neuroscience Center</addr-line>
          <country>UNITED STATES</country>
          <addr-line content-type="city">Chapel Hill</addr-line>
          <addr-line content-type="zipcode">27599</addr-line>
        </aff>
        <aff id="aff2">
          <institution>Xona Microfluidics, LLC</institution>
          <addr-line content-type="addrline1">Xona Microfluidics LLC</addr-line>
          <country>UNITED STATES</country>
          <addr-line content-type="city">Temecula</addr-line>
          <addr-line content-type="state">CA</addr-line>
          <addr-line content-type="zipcode">92590</addr-line>
        </aff>
        <aff id="aff3">
          <institution>Xona Microfluidics, LLC</institution>
          <addr-line content-type="addrline1">Xona Microfluidics LLC</addr-line>
          <country>UNITED STATES</country>
          <addr-line content-type="city">Temecula</addr-line>
          <addr-line content-type="state">CA</addr-line>
          <addr-line content-type="zipcode">92590</addr-line>
        </aff>
        <aff id="aff4">
          <country>UNITED STATES</country>
        </aff>
        <aff id="orcid7">0000-0003-0998-087X</aff>
        <aff id="aff5">
          <institution>JoVE</institution>
          <addr-line>1 Alewife Center</addr-line>
          <addr-line>Suite 200</addr-line>
          <addr-line>Cambridge, MA  02140</addr-line>
          <country>UNITED STATES</country>
        </aff>
      </contrib-group>
      <history>
        <date date-type="received">
          <day>04</day>
          <month>05</month>
          <year>2018</year>
        </date>
        <date date-type="rev-recd">
          <day>20</day>
          <month>07</month>
          <year>2018</year>
        </date>
        <date date-type="accepted">
          <day>24</day>
          <month>07</month>
          <year>2018</year>
        </date>
      </history>
      <abstract>
        <p>Microfabricated methods to compartmentalize neurons have become essential tools for many neuroscientists. This protocol describes the use of a pre-assembled plastic chip (sold commercially as the XonaChip) for compartmentalizing cultured primary rat hippocampal neurons. These plastic chips, contained within the footprint of a standard microscope slide, are compatible with high-resolution, live, and fluorescence imaging. This protocol demonstrates how to retrograde label neurons via isolated axons using a modified rabies virus encoding a fluorescent protein, create isolated microenvironments within one compartment, and perform axotomy and immunocytochemistry on-chip. Neurons are cultured for &gt;3 weeks within the plastic chips, illustrating the compatibility of these chips for long-term neuronal cultures.</p>
      </abstract>
      <kwd-group>
        <kwd>Microfluidic chamber, neuron culture</kwd>
        <kwd>hippocampal neurons</kwd>
        <kwd>XonaChip</kwd>
        <kwd>microfluidic chip</kwd>
        <kwd>cultured neurons</kwd>
        <kwd>compartmentalized chamber</kwd>
        <kwd>primary neurons</kwd>
        <kwd>chip</kwd>
        <kwd>axon isolation</kwd>
      </kwd-group>
      <funding-group />
      <counts>
        <fig-count count="0" />
      </counts>
      <custom-meta-group>
        <custom-meta>
          <meta-name>Please indicate the &lt;b&gt; city, state/province, and country&lt;/b&gt; where this article will be &lt;b&gt;filmed&lt;/b&gt;. Please do not use abbreviations.</meta-name>
          <meta-value>UNC-Chapel Hill, 111 Mason Farm Rd, MBRB room 4304, Chapel Hill, NC 27599</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Black and White Image Count</meta-name>
          <meta-value>0</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Color Image Count</meta-name>
          <meta-value>0</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Production Notes</meta-name>
          <meta-value>look out for commercial names, I fixed the manuscript but the authors may try to reintroduce them during scripting and proofing -alisha</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Acquisitions Editor</meta-name>
          <meta-value>Benjamin Werth</meta-value>
        </custom-meta>
      </custom-meta-group>
    </article-meta>
  </front>
  <body>
    <fig>
      <label>Figures</label>
      <graphic xlink:href="Figures ALL copy.pdf" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <table-wrap>
      <label>Materials Table</label>
      <graphic xlink:href="Copy of JoVE_Materials.xls" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </table-wrap>
    <table-wrap>
      <label>Materials Table</label>
      <graphic xlink:href="" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </table-wrap>
    <fig>
      <label>Figure 2</label>
      <graphic xlink:href="Figures ALL REV copy-02.eps" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 3</label>
      <graphic xlink:href="Figures ALL REV copy-03.eps" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 4</label>
      <graphic xlink:href="Figures ALL REV copy-04.eps" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 5</label>
      <graphic xlink:href="Figures ALL REV copy 05.ai" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 6</label>
      <graphic xlink:href="Figures ALL REV copy-06.eps" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 7</label>
      <graphic xlink:href="Figures ALL REV copy 07.ai" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <table-wrap>
      <label>Table of Materials</label>
      <graphic xlink:href="Copy of JoVE_Materials REV.xls" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </table-wrap>
    <fig>
      <label>Figure 1</label>
      <graphic xlink:href="Figures ALL REV copy-01.pdf" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <table-wrap>
      <label>Table 1</label>
      <graphic xlink:href="table 1- chips vs PDMS chambers.xlsx" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </table-wrap>
  </body>
</article>