{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T22:09:47Z","timestamp":1782338987811,"version":"3.54.5"},"reference-count":22,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2008,1,11]],"date-time":"2008-01-11T00:00:00Z","timestamp":1200009600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cytometry Pt A"],"published-print":{"date-parts":[[2008,2]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The accurate determination of cell cycle, immunophenotypes and morphology at single\u2010cell level is not fully achieved by current flow cytometry protocols. Acetone, a coagulant fixative\/permealizing agent, is widely used in static cytometry, but is impractical in flow cytometry because of its shrinking effect. We sought for conditions of acetone treatment that could permit the simultaneous analysis of physical parameters, surface and intracellular immunostaining, and DNA content. We evaluated different experimental conditions (concentration, duration of fixation, temperature, presence of proteins) to test the capacity of acetone fixation\/permeabilization to preserve cell physical parameters (forward and side scatters, FSC, and SSC) and immunophenotyping while allowing stoichiometric DNA staining. The commonly used ethanol fixation technique was used as reference method. To detect phenotypes and DNA content simultaneously, we employed 7\u2010aminoactinomycin D (7\u2010AAD) as \u201cintercalating\u201d dye for DNA in spite of, or just for, its controversial ability in stoichiometric DNA staining. Cells were resting peripheral blood monucleated cells (PBMCs), T\u2010 and B\u2010cell blasts obtained by PBMCs stimulation, and the human cell lines Ramos and Shep. Acetone fixation, preserving both the recovery and the physical parameters of cells, is drastically influenced by temperature of treatment and is practicable only when the protocol is realized at 8\u00b0C. Under this condition, acetone maintains the immunophenotypic fluorescences (realized before or after the fixation) better than ethanol. Stoichiometric DNA staining of acetone processed cells, the variation coefficients (CV) of frequency distributions of G1\/G0 and G2\/M phases, the modes ratio of these distributions and doublets generation are at least comparable to those obtained with ethanol treatment. The assay developed in the present study, that we called flow acetone\u2010staining technique (FAST), accurately analyzes cell cycle, physical parameters and immunophenotypes in heterogenous cell populations, and thus provides a useful tool for cytomics. \u00a9 2008 International Society for Analytical Cytology.<\/jats:p>","DOI":"10.1002\/cyto.a.20521","type":"journal-article","created":{"date-parts":[[2008,1,11]],"date-time":"2008-01-11T18:49:20Z","timestamp":1200077360000},"page":"168-174","source":"Crossref","is-referenced-by-count":10,"title":["Flow acetone\u2010staining technique: A highly efficient procedure for the simultaneous analysis of DNA content, cell morphology, and immunophenotype by flow cytometry"],"prefix":"10.1002","volume":"73A","author":[{"given":"Maurizio","family":"Carbonari","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Debora","family":"Mancaniello","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tiziana","family":"Tedesco","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Massimo","family":"Fiorilli","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2008,1,11]]},"reference":[{"key":"e_1_2_5_2_2","doi-asserted-by":"publisher","DOI":"10.1023\/A:1002942418520"},{"key":"e_1_2_5_3_2","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.990120312"},{"key":"e_1_2_5_4_2","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.990130109"},{"key":"e_1_2_5_5_2","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.990130414"},{"key":"e_1_2_5_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(94)90373-5"},{"key":"e_1_2_5_7_2","first-page":"8.4.1","volume-title":"Current Protocols in Cell Biology","author":"Darzynkiewicz Z","year":"1999"},{"key":"e_1_2_5_8_2","doi-asserted-by":"publisher","DOI":"10.1177\/32.4.6368682"},{"key":"e_1_2_5_9_2","first-page":"431","article-title":"The amex method. A semplified technique of tissue processing and paraffin embedding with improved preservation of antigens for immunostaining","volume":"125","author":"Sato Y","year":"1986","journal-title":"Am J Pathol"},{"key":"e_1_2_5_10_2","first-page":"507","volume-title":"Practical Immunology","author":"Hudson L","year":"1989"},{"key":"e_1_2_5_11_2","first-page":"270","article-title":"Glucocorticoids and irradiation\u2010induced apoptosis in normal murine bone marrow B\u2010lineage lymphocytes as determined by flow cytometry","volume":"79","author":"Garvy BA","year":"1993","journal-title":"Immunology"},{"key":"e_1_2_5_12_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0898-743"},{"key":"e_1_2_5_13_2","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.136.8.2769"},{"key":"e_1_2_5_14_2","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.164.2.944"},{"key":"e_1_2_5_15_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-4652(200003)182:3<311::AID-JCP1>3.0.CO;2-9"},{"key":"e_1_2_5_16_2","doi-asserted-by":"publisher","DOI":"10.1214\/ss\/1032280305"},{"key":"e_1_2_5_17_2","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1093\/oso\/9780199638253.001.0001","volume-title":"Flow Cytometry","author":"Ormerod MG","year":"2000"},{"key":"e_1_2_5_18_2","doi-asserted-by":"publisher","DOI":"10.1177\/26.11.82573"},{"key":"e_1_2_5_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0091-679X(08)60519-1"},{"key":"e_1_2_5_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-1759(00)00297-0"},{"key":"e_1_2_5_21_2","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.10163"},{"key":"e_1_2_5_22_2","doi-asserted-by":"publisher","DOI":"10.1002\/1097-0320(20010601)44:2<120::AID-CYTO1090>3.0.CO;2-7"},{"key":"e_1_2_5_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2006.06.015"}],"container-title":["Cytometry Part A"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fcyto.a.20521","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cyto.a.20521","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,2,21]],"date-time":"2024-02-21T05:30:47Z","timestamp":1708493447000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/cyto.a.20521"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2008,1,11]]},"references-count":22,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2008,2]]}},"alternative-id":["10.1002\/cyto.a.20521"],"URL":"https:\/\/doi.org\/10.1002\/cyto.a.20521","archive":["Portico"],"relation":{},"ISSN":["1552-4922","1552-4930"],"issn-type":[{"value":"1552-4922","type":"print"},{"value":"1552-4930","type":"electronic"}],"subject":[],"published":{"date-parts":[[2008,1,11]]}}}