{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T23:28:22Z","timestamp":1778714902740,"version":"3.51.4"},"reference-count":29,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2010,1,26]],"date-time":"2010-01-26T00:00:00Z","timestamp":1264464000000},"content-version":"vor","delay-in-days":25,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Contrast Media &amp; Molecular"],"published-print":{"date-parts":[[2010,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p><jats:italic>In vivo<\/jats:italic> optical imaging using fluorescently labeled self\u2010quenched monoclonal antibodies, activated through binding and internalization within target cells, results in excellent target\u2010to\u2010background ratios. We hypothesized that these molecular probes could be utilized to accurately report on cellular internalization with fluorescence lifetime imaging (FLI). Two imaging probes were synthesized, consisting of the antibody trastuzumab (targeting HER2\/neu) conjugated to Alexa Fluor750 in ratios of either 1:8 or 1:1. Fluorescence intensity and lifetime of each conjugate were initially determined at endosomal pHs. Since the 1:8 conjugate is self\u2010quenched, the fluorescence lifetime of each probe was also determined after exposure to the known dequencher SDS. <jats:italic>In vitro<\/jats:italic> imaging experiments were performed using 3T3\/HER2<jats:sup>+<\/jats:sup> and BALB\/3T3 (HER2<jats:sup>\u2212<\/jats:sup>) cell lines. Changes in fluorescence lifetime correlated with temperature\u2010 and time\u2010dependent cellular internalization. <jats:italic>In vivo<\/jats:italic> imaging studies in mice with dual flank tumors [3T3\/HER2<jats:sup>+<\/jats:sup> and BALB\/3T3 (HER2<jats:sup>\u2212<\/jats:sup>)] detected a minimal difference in FLI. In conclusion, fluorescence lifetime imaging monitors the internalization of target\u2010specific activatable antibody\u2013fluorophore conjugates <jats:italic>in vitro<\/jats:italic>. Challenges remain in adapting this methodology to <jats:italic>in vivo<\/jats:italic> imaging. Copyright \u00a9 2010 John Wiley &amp; Sons, Ltd.<\/jats:p>","DOI":"10.1002\/cmmi.360","type":"journal-article","created":{"date-parts":[[2010,1,26]],"date-time":"2010-01-26T09:39:01Z","timestamp":1264498741000},"page":"1-8","source":"Crossref","is-referenced-by-count":23,"title":["Fluorescence lifetime imaging of activatable target specific molecular probes"],"prefix":"10.1002","volume":"5","author":[{"given":"Raphael","family":"Alford","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mikako","family":"Ogawa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Moinuddin","family":"Hassan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amir H.","family":"Gandjbakhche","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter L.","family":"Choyke","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hisataka","family":"Kobayashi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2010,1,26]]},"reference":[{"key":"e_1_2_1_2_2","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1047403"},{"key":"e_1_2_1_3_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.bioeng.8.061505.095831"},{"key":"e_1_2_1_4_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1125949"},{"key":"e_1_2_1_5_2","doi-asserted-by":"publisher","DOI":"10.1021\/mp800115t"},{"key":"e_1_2_1_6_2","doi-asserted-by":"publisher","DOI":"10.1158\/1535-7163.MCT-08-0862"},{"key":"e_1_2_1_7_2","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-08-3116"},{"key":"e_1_2_1_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrc1717"},{"key":"e_1_2_1_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2006.109"},{"key":"e_1_2_1_10_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2006.119"},{"key":"e_1_2_1_11_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0906388106"},{"key":"e_1_2_1_12_2","doi-asserted-by":"publisher","DOI":"10.1007\/b102213"},{"key":"e_1_2_1_13_2","doi-asserted-by":"crossref","first-page":"229","DOI":"10.2310\/7290.2007.00019","article-title":"Fluorescence lifetime imaging system for in vivo studies","volume":"6","author":"Hassan M","year":"2007","journal-title":"Mol Imag"},{"key":"e_1_2_1_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(03)60126-6"},{"key":"e_1_2_1_15_2","doi-asserted-by":"publisher","DOI":"10.1002\/lsm.20101"},{"key":"e_1_2_1_16_2","doi-asserted-by":"publisher","DOI":"10.1023\/B:JOFL.0000039343.02843.12"},{"key":"e_1_2_1_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(92)90112-K"},{"key":"e_1_2_1_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-3495(02)73936-2"},{"key":"e_1_2_1_19_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-008-2147-0"},{"key":"e_1_2_1_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/0143-4160(94)90100-7"},{"key":"e_1_2_1_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/0143-4160(92)90041-P"},{"key":"e_1_2_1_22_2","doi-asserted-by":"publisher","DOI":"10.1007\/s10895-005-2523-5"},{"key":"e_1_2_1_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpa.2003.08.007"},{"key":"e_1_2_1_24_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.onc.1203973"},{"key":"e_1_2_1_25_2","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.98.5.1757"},{"key":"e_1_2_1_26_2","doi-asserted-by":"publisher","DOI":"10.1021\/bc0601626"},{"key":"e_1_2_1_27_2","doi-asserted-by":"publisher","DOI":"10.1158\/1078-0432.CCR-07-1119"},{"key":"e_1_2_1_28_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1349-7006.2009.01133.x"},{"key":"e_1_2_1_29_2","doi-asserted-by":"publisher","DOI":"10.3233\/CBM-2008-4602"},{"key":"e_1_2_1_30_2","doi-asserted-by":"publisher","DOI":"10.1021\/bc8003765"}],"container-title":["Contrast Media &amp; 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