{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T20:41:56Z","timestamp":1790109716672,"version":"4.0.1"},"reference-count":28,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2019,7,13]],"date-time":"2019-07-13T00:00:00Z","timestamp":1562976000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"region Provence-Alpes-C\u00f4te d'azur","award":["emplois jeunes doctorant"],"award-info":[{"award-number":["emplois jeunes doctorant"]}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cytometry Part B Clinical"],"published-print":{"date-parts":[[2019,9]]},"abstract":"<jats:sec><jats:title>BACKGROUND<\/jats:title><jats:p>Flow cytometry essentially focuses on surface\u2010expressed proteins, with few protocols being devoted to intracellular components. We evaluated a two\u2010step procedure using new formaldehyde\u2010free permeabilization and staining reagents that allow the staining of platelets and red blood cells (RBCs) from whole blood.<\/jats:p><\/jats:sec><jats:sec><jats:title>METHODS<\/jats:title><jats:p>Citrated blood was treated with the new staining protocol (NSP) or control reagent (phosphate\u2010buffered solution bovine serum albumin) and stained with antibodies against surface or intracellular markers. The effects of the NSP on cell integrity, morphology, and content were evaluated.<\/jats:p><\/jats:sec><jats:sec><jats:title>RESULTS<\/jats:title><jats:p>The NSP slightly reduced the cell count (~20%) and changed the RBC morphology with a 42% mean diameter reduction. Conversely, the NSP did not affect platelet discoid morphology and led to a minor size decrease (11%). These morphological changes neither impelled a gating strategy modification nor interfered with the discrimination among populations based on surface markers. The NSP provided intracellular access to all the tested antigens: CD62P, FXIII, and CD63 in platelets and glycated and fetal hemoglobin (HbA1c and HbF) and nucleic acid in RBCs. The NSP gave excellent intra\u2010assay precision with minimal impact on cell morphology and fluorescence labelling over time (up to 24\u2009h).<\/jats:p><\/jats:sec><jats:sec><jats:title>CONCLUSIONS<\/jats:title><jats:p>With the ability to detect surface and intracellular antigens through a rapid preparation protocol without washing steps or toxic formaldehyde treatment, this NSP designed for research offers a marked improvement in the analysis of platelets and RBCs isolated directly from whole blood. Consequently, the NSP opens new avenues to investigate platelet degranulation and erythrocyte subpopulations. \u00a9 2019 International Clinical Cytometry Society<\/jats:p><\/jats:sec>","DOI":"10.1002\/cyto.b.21839","type":"journal-article","created":{"date-parts":[[2019,7,13]],"date-time":"2019-07-13T09:51:00Z","timestamp":1563011460000},"page":"426-435","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["A Novel Rapid Method of Red Blood Cell and Platelet Permeabilization and Staining for Flow Cytometry Analysis"],"prefix":"10.1002","volume":"96","author":[{"given":"Sixtine","family":"Faur\u00e9","sequence":"first","affiliation":[{"name":"Aix\u2010Marseille Univ INSERM, INRA, C2VN  Marseille France"},{"name":"Beckman Coulter Life Sciences Immunotech  Marseille France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Van 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