{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T19:37:11Z","timestamp":1790797031726,"version":"4.1.0"},"reference-count":29,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2019,12,23]],"date-time":"2019-12-23T00:00:00Z","timestamp":1577059200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"National Cancer Institute to the Roswell Park Comprehensive Cancer Center","award":["2 P30 CA016056-42"],"award-info":[{"award-number":["2 P30 CA016056-42"]}]},{"name":"National Institutes of Health's Shared Instrument Program"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cytometry Part B Clinical"],"published-print":{"date-parts":[[2020,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Recent advances in therapeutic interventions have dramatically improved complete response rates in patients with multiple myeloma (MM). The ability to identify residual myeloma cells (e.g., measurable residual disease [MRD]) can provide valuable information pertaining to patient's depth of response to therapy and risk of relapse. Multiparametric flow cytometry is an excellent technique to monitor MRD and has been demonstrated to correlate with patient outcome post\u2010treatment. To achieve the high sensitivity (one abnormal cell in 10<jats:sup>5<\/jats:sup>\u201310<jats:sup>6<\/jats:sup> cells) required for MRD evaluation, millions of cells have to be acquired and conventional immunophenotyping protocols are unable to attain these numbers, indicating the needs for alternative flow cytometric staining procedures. A bulk, \u201cPre\u2010lysis\u201d method is the consensus approach for staining large number of cells, requires two red blood cell lysis steps, and can adversely affect epitope density. In this study, we tested the \u201cPooled\u2010tube\u201d and \u201cDextran Sedimentation\u201d staining procedures and correlated them with the \u201cPre\u2010lysis\u201d method as potential alternative approaches.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>A total of 22 bone marrow aspirates from patients with plasma cell (PC) dyscrasia were processed in parallel using the \u201cPre\u2010lysis,\u201d \u201cPooled\u2010tube,\u201d and \u201cDextran Sedimentation\u201d techniques. Stain indices were calculated and compared to assess their impacts on staining performance for each antibody used in the consensus panel. The recovery of normal and abnormal PCs, mast cells, and B cell precursors was enumerated and compared after their counts were normalized using fluorescent beads. The limit of blank, limit of detection, and lower limit of quantification were established using serial dilution experiments.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The staining performances of CD19 PECy7, CD27 BV510, CD81 APCH7, and CD138 BV421 were improved using the \u201cPooled\u2010tube\u201d method when compared to \u201cPre\u2010lysis.\u201d \u201cPre\u2010lysis\u201d was better at resolving CD56 using clone C5.9 but our results demonstrated similar improvement can also be achieved by \u201cPooled\u2010tube\u201d when alternative CD56 PE clones were used. \u201cDextran sedimentation\u201d yielded similar staining results when compared to \u201cPre\u2010lysis\u201d for all the markers analyzed. The \u201cPooled\u2010tube\u201d method, when normalized to \u201cPre\u2010lysis,\u201d recovered higher numbers of total PCs (1.2\u2009\u00b1\u20090.2 times higher; <jats:italic>p<\/jats:italic> = .049), normal PCs (1.4\u2009\u00b1\u20090.26; <jats:italic>p<\/jats:italic> = .007), mast cells (1.46\u2009\u00b1\u20090.27; <jats:italic>p<\/jats:italic> = .003), and B cell precursors (1.42\u2009\u00b1\u20090.3; <jats:italic>p<\/jats:italic> = .011), but not abnormal PCs (1.09\u2009\u00b1\u20090.2; <jats:italic>p<\/jats:italic> = .352). There was no evidence that the recovery of cells was different between \u201cPre\u2010lysis\u201d versus \u201cDextran Sedimentation.\u201d All three flow cytometric assays achieved a minimum sensitivity of 10<jats:sup>\u22125<\/jats:sup> and approached that of 10<jats:sup>\u22126<\/jats:sup> for detecting rare events.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Both \u201cPooled\u2010tube\u201d and \u201cDextran Sedimentation\u201d staining procedures were comparable to the \u201cPre\u2010lysis\u201d method and are suitable high sensitivity flow cytometric approaches that can be used to process bone marrow samples for MM MRD testing.<\/jats:p><\/jats:sec>","DOI":"10.1002\/cyto.b.21862","type":"journal-article","created":{"date-parts":[[2019,12,23]],"date-time":"2019-12-23T11:57:05Z","timestamp":1577102225000},"page":"161-173","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":30,"title":["Methodological considerations for the high sensitivity detection of multiple myeloma measurable residual disease"],"prefix":"10.1002","volume":"98","author":[{"given":"Kah Teong","family":"Soh","sequence":"first","affiliation":[{"name":"Department of Flow and Image Cytometry Roswell Park Comprehensive Cancer Center  Buffalo New York"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"suffix":"Jr.","given":"Joseph D.","family":"Tario","sequence":"additional","affiliation":[{"name":"Department of Flow and Image Cytometry Roswell Park Comprehensive Cancer Center  Buffalo New York"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Theresa E","family":"Hahn","sequence":"additional","affiliation":[{"name":"Department of Medicine Roswell Park Comprehensive Cancer Center  Buffalo New York"},{"name":"Transplant and Cellular Therapy Program Roswell Park Comprehensive Cancer Center  Buffalo New York"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jens","family":"Hillengass","sequence":"additional","affiliation":[{"name":"Department of Medicine Roswell Park Comprehensive Cancer Center  Buffalo New York"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philip L.","family":"McCarthy","sequence":"additional","affiliation":[{"name":"Department of Medicine Roswell Park Comprehensive Cancer Center  Buffalo New York"},{"name":"Transplant and Cellular Therapy Program Roswell Park Comprehensive Cancer Center  Buffalo New York"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul K.","family":"Wallace","sequence":"additional","affiliation":[{"name":"Department of Flow and Image Cytometry Roswell Park Comprehensive Cancer Center  Buffalo New York"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2019,12,23]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.clml.2018.06.015"},{"issue":"1","key":"e_1_2_6_3_1","first-page":"S49","article-title":"Limit of blank, limit of detection and limit of quantitation","volume":"29","author":"Armbruster D. 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