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Current quantification methods of the engulfment of apoptotic cells by macrophages are potentially flawed by several limitations. Adherent macrophage populations are overlaid with apoptotic targets in suspension and then co\u2010cultured for a definite period, which may give rise to two different features: (1) engulfed and (2) non\u2010engulfed macrophages that are surface\u2010bound cell populations. Rigorous washing to dislodge surface\u2010bound apoptotic cells before assessment of phagocytosis may lead to loss of phagocytes, thereby skewing the apparent magnitude of the overall phagocytic response. There is a need for simple and reliable methods to clearly determine the internalization of apoptotic cells. In this unit, we demonstrate the use of pHrodo\u2010succinimidyl ester (SE), a pH\u2010sensitive fluorescent dye, to label the apoptotic cells for monitoring the phagocytosis. After engulfment, the intensity of pHrodo light emission will be elevated due to the pH change inside of macrophages. The shift of pHrodo light emission can be detected by a flow cytometer or using a fluorescence microscope. <jats:italic>Curr. Protoc. Immunol<\/jats:italic>. 100:14.31.1\u201014.31.8. \u00a9 2013 by John Wiley &amp; Sons, Inc.<\/jats:p>","DOI":"10.1002\/0471142735.im1431s100","type":"journal-article","created":{"date-parts":[[2013,2,7]],"date-time":"2013-02-07T19:33:01Z","timestamp":1360265581000},"source":"Crossref","is-referenced-by-count":16,"title":["Measurement of Phagocytic Engulfment of Apoptotic Cells by Macrophages Using pHrodo Succinimidyl Ester"],"prefix":"10.1002","volume":"100","author":[{"given":"Monowar","family":"Aziz","sequence":"first","affiliation":[{"name":"Center for Immunology and Inflammation, The Feinstein Institute for Medical Research and Department of Surgery, Hofstra North Shore\u2010LIJ School of Medicine  Manhasset New York"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weng\u2010Lang","family":"Yang","sequence":"additional","affiliation":[{"name":"Center for Immunology and Inflammation, The Feinstein Institute for Medical Research and Department of Surgery, Hofstra North Shore\u2010LIJ School of Medicine  Manhasset New York"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"Wang","sequence":"additional","affiliation":[{"name":"Center for Immunology and Inflammation, The Feinstein Institute for Medical Research and Department of Surgery, Hofstra North Shore\u2010LIJ School of Medicine  Manhasset New York"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2013,2]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/35046549"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.148.7.2207"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/417182a"},{"key":"e_1_2_6_5_1","first-page":"162","article-title":"MFG\u2010E8\u2010dependent clearance of apoptotic cells, and autoimmunity caused by its failure","volume":"9","author":"Hanayama R.","year":"2006","journal-title":"Curr. 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