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<!DOCTYPE article SYSTEM "archivearticle3.dtd">
<article dtd-version="3.0" article-type="Invited Methods Article - JoVE Produced Video">
  <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">JoVE58509</article-id>
      <article-id pub-id-type="manuscript">JoVE58509</article-id>
      <article-categories>
        <subj-group subj-group-type="Article Type">
          <subject>Invited Methods Article - JoVE Produced Video</subject>
        </subj-group>
        <subj-group subj-group-type="Category">
          <subject>JoVE Immunology and Infection</subject>
        </subj-group>
        <subj-group subj-group-type="Classification">
          <subject>Pathologic Processes</subject>
          <subject>Inflammation</subject>
          <subject>Inflammation Mediators</subject>
          <subject>immunology</subject>
          <subject>Diseases</subject>
          <subject>Pathological Conditions, Signs and Symptoms</subject>
          <subject>Chemicals and Drugs</subject>
          <subject>Biological Factors</subject>
          <subject>Life sciences</subject>
          <subject>Life Sciences (General)</subject>
        </subj-group>
      </article-categories>
      <title-group>
        <article-title>Simultaneous study of the recruitment of monocyte subpopulations under flow in vitro</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author">
          <role content-type="1" />
          <name>
            <surname>Ropraz</surname>
            <given-names>Patricia</given-names>
          </name>
          <email>Patricia.Ropraz@unige.ch</email>
          <xref ref-type="aff" rid="aff1" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="2" />
          <name>
            <surname>Imhof</surname>
            <given-names>Beat  A</given-names>
          </name>
          <email>Beat.Imhof@unige.ch</email>
          <xref ref-type="aff" rid="aff2" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="3" />
          <name>
            <surname>Matthes</surname>
            <given-names>Thomas</given-names>
          </name>
          <email>Thomas.Matthes@unige.ch</email>
          <xref ref-type="aff" rid="aff3" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="4" />
          <name>
            <surname>Wehrle-Haller</surname>
            <given-names>Bernhard</given-names>
          </name>
          <email>Bernhard.Wehrle-Haller@unige.ch</email>
          <xref ref-type="aff" rid="aff4" />
        </contrib>
        <contrib contrib-type="author" corresp="yes">
          <role content-type="5" />
          <name>
            <surname>Sidibe</surname>
            <given-names>Adama</given-names>
            <prefix>Dr.</prefix>
          </name>
          <email>Adama.Sidibe@unige.ch</email>
          <xref ref-type="aff" rid="aff5" />
        </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="aff6" />
        </contrib>
        <aff id="aff1">
          <institution>Universite de Geneve</institution>
          <institution content-type="dept">Hematology Service</institution>
          <addr-line content-type="addrline1">Michel-Servet 1</addr-line>
          <country>SWITZERLAND</country>
          <addr-line content-type="city">Genève</addr-line>
          <addr-line content-type="zipcode">1211</addr-line>
        </aff>
        <aff id="aff2">
          <institution>Universite de Geneve</institution>
          <institution content-type="dept">Department of Pathology and Immunology</institution>
          <addr-line content-type="addrline1">Michel-Servet 1</addr-line>
          <country>SWITZERLAND</country>
          <addr-line content-type="city">Genève</addr-line>
          <addr-line content-type="zipcode">1211</addr-line>
        </aff>
        <aff id="aff3">
          <institution>Universite de Geneve</institution>
          <institution content-type="dept">Hematology Service</institution>
          <addr-line content-type="addrline1">Michel-Servet 1</addr-line>
          <country>SWITZERLAND</country>
          <addr-line content-type="city">Genève</addr-line>
          <addr-line content-type="zipcode">1211</addr-line>
        </aff>
        <aff id="aff4">
          <institution>Universite de Geneve</institution>
          <institution content-type="dept">Department of Cell Physiology and Metabolism</institution>
          <addr-line content-type="addrline1">Michel-Servet 1</addr-line>
          <country>SWITZERLAND</country>
          <addr-line content-type="city">Genève</addr-line>
          <addr-line content-type="zipcode">1211</addr-line>
        </aff>
        <aff id="aff5">
          <institution>Universite de Geneve</institution>
          <institution content-type="dept">Department of Cell Physiology and Metabolism</institution>
          <addr-line content-type="addrline1">Michel-Servet 1</addr-line>
          <country>SWITZERLAND</country>
          <addr-line content-type="city">Genève</addr-line>
          <addr-line content-type="zipcode">1211</addr-line>
        </aff>
        <aff id="aff6">
          <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>23</day>
          <month>05</month>
          <year>2018</year>
        </date>
        <date date-type="rev-recd">
          <day>24</day>
          <month>08</month>
          <year>2018</year>
        </date>
        <date date-type="rev-recd">
          <day>07</day>
          <month>08</month>
          <year>2018</year>
        </date>
        <date date-type="accepted">
          <day>29</day>
          <month>08</month>
          <year>2018</year>
        </date>
      </history>
      <abstract>
        <p>The recruitment of monocytes from the blood to targeted peripheral tissues is critical to the inflammatory process during tissue injury, tumor development and autoimmune diseases. This is facilitated through a process of capture from free flow onto the luminal surface of activated endothelial cells, followed by their adhesion and transendothelial migration (transmigration) into the underlying affected tissue. However, the mechanisms that support the preferential and context-dependent recruitment of monocyte subpopulations are still not fully understood. Therefore, we have developed a method that allows the recruitment of different monocyte subpopulations to be simultaneously visualized and measured under flow. This method, based on time-lapse confocal imaging, allows for the unambiguous distinction between adherent and transmigrated monocytes. Here, we describe how this method can be used to simultaneously study the recruitment cascade of pro-angiogenic and non-angiogenic monocytes in vitro. Furthermore, this method can be extended to study the different steps of recruitment of up to three monocyte populations.</p>
      </abstract>
      <kwd-group>
        <kwd>Leukocyte trafficking | Monocyte recruitment | Inflammation | Timelapse imaging | Confocal microscopy | Adhesion | Transmigration | Monocyte subpopulations | Chemokines | Integrins | Adhesion molecules</kwd>
      </kwd-group>
      <funding-group />
      <counts>
        <fig-count count="0" />
      </counts>
      <custom-meta-group>
        <custom-meta>
          <meta-name>Please indicate whether this article will be Standard Access or Open Access. </meta-name>
          <meta-value>Standard Access (US$2,400);</meta-value>
        </custom-meta>
        <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>Buildng D, 6th floor, Department of Cell Physiology and Metabolism, Centre Medical Universitaire, Université de Genève, Michel-Servet 1, 1211 Geneva, Switzerland</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>Acquisitions Editor</meta-name>
          <meta-value>indrani mukherjee</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Assistant who entered lead</meta-name>
          <meta-value>kaitlin cleary</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>ALA Access Type</meta-name>
          <meta-value>JPV Standard Access ($2400)</meta-value>
        </custom-meta>
      </custom-meta-group>
    </article-meta>
  </front>
  <body>
    <table-wrap>
      <label>Materials Table</label>
      <graphic xlink:href="Sidibe-JoVE_Materials.xls" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </table-wrap>
    <fig>
      <label>Supplemental Movie 1</label>
      <graphic xlink:href="Movie1_Apical_.avi" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Supplemental Movie 2</label>
      <graphic xlink:href="Movie2_Basal_.avi" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Supplemental Movie 7</label>
      <graphic xlink:href="Movie7_Basal_TNFVEGF_PMo.avi" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Supplemental Movie 6</label>
      <graphic xlink:href="Movie6_Apical_TNFVEGF_PMo.avi" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Supplemental Movie 3</label>
      <graphic xlink:href="Movie3_Whole_MAX.avi" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Supplemental Movie 4</label>
      <graphic xlink:href="Movie4_Apical_TNF_PMo.avi" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Supplemental Movie 5</label>
      <graphic xlink:href="Movie5_Basal_TNF_PMo.avi" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 1</label>
      <graphic xlink:href="Fig1_PBMC and Mono populations.png" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 2</label>
      <graphic xlink:href="FIG2-Tubing-setting-Flow.png" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 3</label>
      <graphic xlink:href="Fig3_EC shape.png" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 4</label>
      <graphic xlink:href="Fig4_EC_discontinuation.png" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 5</label>
      <graphic xlink:href="Fig5_ECstimulation_PMo TEM.png" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 6</label>
      <graphic xlink:href="Fig6_PBMC_CD14vsCD14neg.png" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <table-wrap>
      <label>Table of Materials</label>
      <graphic xlink:href="JoVE_Materials.xls" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </table-wrap>
    <fig>
      <label>Figure 1</label>
      <graphic xlink:href="Fig1_PBMC and Mono populations.eps" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 2</label>
      <graphic xlink:href="Fig2 Tubing setting Flow.eps" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 3</label>
      <graphic xlink:href="Fig3_EC shape.eps" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 4</label>
      <graphic xlink:href="Fig4_EC_discontinuation.eps" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 5</label>
      <graphic xlink:href="Fig5_ECstimulation_PMo TEM.eps" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 6</label>
      <graphic xlink:href="Fig6_PBMC_CD14vsCD14neg.eps" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <table-wrap>
      <label>Table of Materials</label>
      <graphic xlink:href="JoVE_Materials.xlsx" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </table-wrap>
  </body>
</article>