{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,12]],"date-time":"2026-04-12T02:10:40Z","timestamp":1775959840800,"version":"3.50.1"},"reference-count":7,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2014,8,6]],"date-time":"2014-08-06T00:00:00Z","timestamp":1407283200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Histopathology"],"published-print":{"date-parts":[[2014,11]]},"abstract":"<jats:sec><jats:title>Aims<\/jats:title><jats:p>Current immunohistochemical methods to study the expression of multiple proteins in a single tissue section suffer from several limitations. In this article, we report on sequential immunohistochemistry (S\u2010<jats:styled-content style=\"fixed-case\">IHC<\/jats:styled-content>), a novel, easy method that allows the study of numerous proteins in a single tissue section, while requiring very limited optimization.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods and results<\/jats:title><jats:p>In S\u2010<jats:styled-content style=\"fixed-case\">IHC<\/jats:styled-content>, a tissue section is stained for multiple antibodies, with intermediate scanning of the section and elution of chromogen and antibodies. Overlays are made of the digital images, allowing assessment of multiple proteins in the same tissue section. We used S\u2010<jats:styled-content style=\"fixed-case\">IHC<\/jats:styled-content> to study nine nodular lymphocyte\u2010predominant Hodgkin lymphomas (<jats:styled-content style=\"fixed-case\">NLPHL<\/jats:styled-content>s) and 10 T\u2010cell\u2010rich and histiocyte\u2010rich diffuse large B\u2010cell lymphomas (T\/<jats:styled-content style=\"fixed-case\">HRBCL<\/jats:styled-content>s) for expression of cyclin D1, <jats:styled-content style=\"fixed-case\">CD<\/jats:styled-content>20, and <jats:styled-content style=\"fixed-case\">CD<\/jats:styled-content>68. We observed cyclin D1 expression in single tumour cells in 44% of <jats:styled-content style=\"fixed-case\">NLPHL<\/jats:styled-content>s and 60% of T\/<jats:styled-content style=\"fixed-case\">HRBCL<\/jats:styled-content>s. Comparison of S\u2010<jats:styled-content style=\"fixed-case\">IHC<\/jats:styled-content> with classic single immunohistochemical staining revealed discrepancies in eight cases (42%), demonstrating the difficulty of differentiating tumour cells from histiocytes on morphological grounds, and stressing the additional value of S\u2010<jats:styled-content style=\"fixed-case\">IHC<\/jats:styled-content>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>For research and diagnostic purposes, S\u2010<jats:styled-content style=\"fixed-case\">IHC<\/jats:styled-content> is a promising technique that assesses the expression of numerous proteins in single tissue sections with complete architectural information, allowing phenotypic characterization of single cells.<\/jats:p><\/jats:sec>","DOI":"10.1111\/his.12446","type":"journal-article","created":{"date-parts":[[2014,4,26]],"date-time":"2014-04-26T06:48:39Z","timestamp":1398494919000},"page":"651-657","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":42,"title":["Sequential immunohistochemistry: a promising new tool for the pathology laboratory"],"prefix":"10.1111","volume":"65","author":[{"given":"Michiel","family":"van den Brand","sequence":"first","affiliation":[{"name":"Department of Pathology Radboud University Medical Centre Nijmegen The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brigiet M","family":"Hoevenaars","sequence":"additional","affiliation":[{"name":"Laboratory of Pathology PAMM Laboratories Eindhoven The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jessica H M","family":"Sigmans","sequence":"additional","affiliation":[{"name":"Department of Pathology Radboud University Medical Centre Nijmegen The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jos W R","family":"Meijer","sequence":"additional","affiliation":[{"name":"Department of Pathology Rijnstate Hospital Arnhem The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Patricia H J","family":"van Cleef","sequence":"additional","affiliation":[{"name":"Department of Pathology Radboud University Medical Centre Nijmegen The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Patricia J T A","family":"Groenen","sequence":"additional","affiliation":[{"name":"Department of Pathology Radboud University Medical Centre Nijmegen The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Konnie M","family":"Hebeda","sequence":"additional","affiliation":[{"name":"Department of Pathology Radboud University Medical Centre Nijmegen The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J Han J M","family":"van Krieken","sequence":"additional","affiliation":[{"name":"Department of Pathology Radboud University Medical Centre Nijmegen The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2014,8,6]]},"reference":[{"key":"e_1_2_7_2_1","volume-title":"WHO classification of tumours of haematopoietic and lymphoid tissues","author":"Swerdlow SH","year":"2008"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0032043"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.2353\/jmoldx.2006.050083"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1369\/jhc.2009.953612"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1177\/26.4.207771"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1369\/jhc.2009.953240"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2559.2011.04104.x"}],"container-title":["Histopathology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fhis.12446","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/his.12446","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,3]],"date-time":"2023-10-03T06:18:05Z","timestamp":1696313885000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/his.12446"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,8,6]]},"references-count":7,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2014,11]]}},"alternative-id":["10.1111\/his.12446"],"URL":"https:\/\/doi.org\/10.1111\/his.12446","archive":["Portico"],"relation":{},"ISSN":["0309-0167","1365-2559"],"issn-type":[{"value":"0309-0167","type":"print"},{"value":"1365-2559","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,8,6]]},"assertion":[{"value":"2014-01-20","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2014-04-23","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2014-08-06","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}