Clinical Cytometry and Immunophenotyping

Patrick, Catherine W.; Ash, Robert C.; Libnoch, Joseph A.; Keller, R.

doi:10.1159/000157042

Cited by 10 publications

(2 citation statements)

References 17 publications

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“…To exclude unspecific staining of lymphocytes, B-lymphocyte analysis is improved when CD19 is analyzed in combination with forward and sideward scattering discrimination, shown in Fig. 1 (27,33,35,44). In the present study PE-or PE-Cy5-conjugated antibodies were used for the low antigen density of CD5 and CD19, whereas a PerCP-conjugated monoclonal antibody was used for the medium to high antigen density of CD20.…”

Section: Discussionmentioning

confidence: 99%

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Multiparametric immunophenotyping of B cells in peripheral blood of healthy adults by flow cytometry

Höffkes

Schmidtke

Uppenkamp

et al. 1996

Clin Diagn Lab Immunol

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The investigation of patients suffering from malignant lymphomas of the B-cell type requires flow cytometric immunophenotyping. Several reports described the expression of almost all B lineage antigens on normal and abnormal B lymphocytes. Thus, immunophenotyping of lymphomas must be interpreted in the context of the reference values obtained for healthy control individuals. For this purpose multiparametric flow cytometric analysis offers the unique feature for lymphocyte subset analysis. In the present study B lymphocytes in the peripheral blood of healthy adults were investigated by multiparametric flow cytometric immunophenotyping for the detection of the frequency (in percent) of antigens provided by the revised European-American classification of lymphoid neoplasms (REAL) classification. Thus, 84 healthy adults were investigated and grouped by age (average ages were as follows: group 1, 25.38 years; group 2, 33.86 years; group 3, 44.17 years; group 4, 55.67 years; group 5, 66.67 years). Analysis was done for surface immunoglobulins (kappa and lambda chains of immunoglobulin M [IgM] and IgD) as well as CD10, CD11c, CD23, CD38, CD103, FMC-7, and B-B4. Three-color immunophenotyping was performed for kappa/CD19/CD5, lambda/CD19/CD5, surface IgM/surface IgD/CD19, FMC-7/CD19/CD5, CD103/CD11c/CD19, CD10/CD23/CD19, and CD38/B-B4/CD19 by live gating of CD19+ events (n = 2,000). Although some numerical differences could be obtained for the different groups, statistical differences (P < 0.005) could only be obtained for the CD19+/CD5+ B-cell subset, which was decreased in the elderly patients (group 5). The established two-color and three-color stainings will serve as a basis for future multiparametric immunophenotyping of abnormal lymphocytes (e.g., for patients suffering from non-Hodgkin's lymphoma of the B-cell type).

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Section: Discussionmentioning

confidence: 99%

“…This approach has been performed with limited marker combinations (43,45,46,48) by using one-color staining or two-color stainings (43,44). In addition, most studies have used lymphocytes prepared by density gradient centrifugation (32,33,39,40). However, preparation by erythrocyte lysis is preferred for flow cytometric immunophenotyping (11,13,31,41).…”

mentioning

confidence: 99%

Multiparametric immunophenotyping of B cells in peripheral blood of healthy adults by flow cytometry

Höffkes

Schmidtke

Uppenkamp

et al. 1996

Clin Diagn Lab Immunol

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Differential analysis of animal bone marrow by flow cytometry

Martin¹,

Brott²,

Zandee³

et al. 1992

Cytometry

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A simple procedure was developed for rapid analysis of animal bone marrow by flow cytometry using the lipophilic cationic dye 3,3′‐dihexyloxacarbocyanine iodide (DiOC6(3)). The batch process allows differentiation of bone marrow cells into lymphoid, erythroid, and myeloid populations and enables classification of erythroid and myeloid cells into proliferating and maturing subpopulations. From these data, myeloid:erythroid (M:E) ratios and maturation indices for erythroid and myeloid cells (EMI and MMI, respectively) can be derived. This procedure provides the opportunity to analyze bone marrow quantitatively and offers distinct advantages to current manual methods in terms of simplicity, throughput, and reproducibility. The method has been tested successfully using marrow from Wistar rats, B6C3F1 mice, beagle dogs, and cynomolgus monkeys. This technique facilitates the evaluation of bone marrow samples taken from preclinical safety studies or from animal colonies of large size. © 1992 Wiley‐Liss, Inc.

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Quality control in flow cytometry for diagnostic pathology: II. A conspectus of reference ranges for lymphocyte immunophenotyping

McCoy

Overton

1994

Cytometry

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Immunophenotyping, as many other clinical assays, is interpreted only in the context of reference values obtained from healthy control individuals. While the use of these reference values, or ranges, has been commonplace in the clinical flow cytometry laboratory for well over a decade, there has been little consensus in standardizing how these values should be obtained, analyzed, or expressed. This report reviews the variables to be considered in establishing reference ranges and statistical methods which can be used. Additionally, examples are given of previously published reference ranges for a variety of specimens often submitted for immunophenotyping. D

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Clinical Cytometry and Immunophenotyping

Cited by 10 publications

References 17 publications

Multiparametric immunophenotyping of B cells in peripheral blood of healthy adults by flow cytometry

Multiparametric immunophenotyping of B cells in peripheral blood of healthy adults by flow cytometry

Differential analysis of animal bone marrow by flow cytometry

Quality control in flow cytometry for diagnostic pathology: II. A conspectus of reference ranges for lymphocyte immunophenotyping

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