Mature B-cell lymphoproliferative disease in patients >30yrs was screened by monoclonal antibodies mixed with CD45-V500c, CD19-Pacific Blue, CD20-APC, CD38-PerCP-Cy5.5, CD10-PE-Cy7, kappa-FITC, and lambda-PE. B-cell and plasma cells in patients >30yrs were screened with a tube containing CD45-V500c, CD19-Pacific Blue, CD20-APC-Cy7, CD38-PerCP-Cy5.5, CD56-ECD, CD138-APC, cytoplasmic κ-FITC, and cytoplasmic λ-PE. The samples were all stained with these T- and B-cell lymphatic screening tubes.
This article uses a method of MFC sample staining and panel screening (14 (link)). If the screening tube detected that the B lymphocytes had an abnormal immunophenotype, we continued to label fluorescent antibody Ki67; Kappa-FITC/Lambda-PE were purchased from DAKO; CD19-Pacific Blue, and CD20-Pacific Blue were purchased from Biolegend. The BD Pharmingen FITC Mouseanti-Ki67 Set (Clone: B56) is compatible with the BD IntraSure™ Kit (641776), and the remainder was purchased from the Becton Dickinson Company (BD, San Jose, CA). Intracellular staining with Ki67 was performed according to the manufacturer’s instructions. At least 5000 abnormal CDl9+ B lymphocytes or a total of 3×105 white blood cells were obtained from each tube. The abnormal B lymphocyte population was gated by CD45/SSC, CDl9/SSC, CDl9/FSC, CD20/SSC, and CDl9/CD20, and the positive rate of Ki67 in these cells was analyzed. The gating strategy for detecting the positive rate of Ki67 expression in non-Hodgkin B-cell lymphoma by flow cytometry is shown in theSupplementary Figure
This article uses a method of MFC sample staining and panel screening (14 (link)). If the screening tube detected that the B lymphocytes had an abnormal immunophenotype, we continued to label fluorescent antibody Ki67; Kappa-FITC/Lambda-PE were purchased from DAKO; CD19-Pacific Blue, and CD20-Pacific Blue were purchased from Biolegend. The BD Pharmingen FITC Mouseanti-Ki67 Set (Clone: B56) is compatible with the BD IntraSure™ Kit (641776), and the remainder was purchased from the Becton Dickinson Company (BD, San Jose, CA). Intracellular staining with Ki67 was performed according to the manufacturer’s instructions. At least 5000 abnormal CDl9+ B lymphocytes or a total of 3×105 white blood cells were obtained from each tube. The abnormal B lymphocyte population was gated by CD45/SSC, CDl9/SSC, CDl9/FSC, CD20/SSC, and CDl9/CD20, and the positive rate of Ki67 in these cells was analyzed. The gating strategy for detecting the positive rate of Ki67 expression in non-Hodgkin B-cell lymphoma by flow cytometry is shown in the
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