3.4.1. Interpretative Guidelines
1. Examine CD45 fluorescence patterns on each specimen histogram. Problems with lysis should not affect results as the CD45-gating strategy focuses on leukocyte populations as positive and other "noise" as negative.
2. Note any abnormal scattering characteristics for the particular panel (i.e., dim CD45 populations).
3. Note any abnormal marking characteristics that may not be appropriate for the particular population selected or which may indicate additional testing to aid in diagnostic interpretation.
4. Always correlate flow cytometric results with morphology review. Any discrepancies should be investigated.
Record percent positive (include double marking) with respect to specific markers tested. Each completed specimen protocol sheet with history, viability, differential, fluorescent marking characteristics, and related histograms should be reviewed. Positivity and negativity for certain markers are characteristics for specific differentiation and growth of hematopoietic cells. Specific diagnosis can be rendered based on these characteristics. The following is an algorithm that can be used for the differential diagnosis of various hematopoietic neoplasma. However, final diagnosis should be based on integrating information from clinical history, histological and morphological evaluation, other laboratory data, and clinical history (1,6,8-10,12-19).
1 Bauer, K. D., Duque, R. E., and Shankey, T. V. (1993) Clinical Flow Cytometry: Principles and Applications, Williams and Wilkins Baltimore, MD USA.
2 Bowman, G. P., Neame, P. B., and Soambonsrup, P. (1986) The contribution of cytochemistry and immunophenotyping to the reproducibility of the FAB classification in acute leukemia. Blood 68, 900-905.
3 Falini, B., Pileri, S., and Martelli, M. F. (1989) Histologic and immunohistological analysis of human lymphomas. Crit. Rev. Oncol. Hematol. 9, 351-419.
4 Harrington, D. S., Masih, A., and Duggan, M. (1991) Immunohistochemical diagnosis of lymphoproliferative diseases. Crit. Rev. Oncol. Hematol. 11, 137-164.
5 Borowitz, M., Duque, R., Bauer, K., et al. (1998) Clinical Applications of Flow Cytometry: Quality Assurance and Immunophenotyping of Lymphocytes. National Committe for Clinical Laboratory Standards.
6 Keren, D. F. (2005) Flow Cytometry in Clinical Diagnosis. ASCP, Chicago, IL.
7 National Committe for Clinical Laboratory Standards document H43-A. (1998) Clinical Applications of Flow Cytometry: Immunophenotyping of Leukemic Cells; Proposed Guideline. National Committee for Clinical Laboratory Standards.
8 Sobol, R.E., Bloomfield, C.D., and Royston, I. (1988) Immunophenotyping in the diagnosis and classification of Acute Lymphoblastic Leukemia. In Clinics in Laboratory Medicine; classification, diagnosis, and molecular biology of Lymphoproliferative disorder. Vol 8, 151-162. W. B. Saunders Company, Philadelphia, PA.
9 Nadler, L. M. and Freedman, A. S. (1987) Cell surface markers in hematologic malignancies. Semin. Oncol. 14, 193-212.
10 Ricker, L. J., Weiss, L. M., Medeiros, L. J., Wood, G. S., and Warnke, R. A. (1987) Immunophenotypic criteria for the diagnosis of non-Hodgkin's lymphoma. Am. J. Pathol. 128, 181-201.
11 Braylan, R. C., Orfao, A., Borowitz, M. J., et al. (2001) Optimal number of reagents required to evaluate hematolymphoid neoplasias: results of an international consensus meeting. Cytometry 46, 23-27.
12 Shapiro, H. M. (2003) Practical Flowcytometry. Wiley-Liss, New York.
13 Foon, K. A., and Todd, R. R. III. (1986) Immunologic classification of leukemia and lymphoma. Blood 68, 1-31.
14 Griffin, J. D., Mayer, R. J., Weinstern, J. J., et al. (1983) Surface marker analysis of acute (myeloblastic leukemia) identification of differentiation-associated pheno-types. Blood 62, 557-563
15 Nguyen, D. T., Diamond, L. W., and Braylan, R. C. (2003) Flow Cytometry in Hematopathology: A Visual Approach to Data Analysis and Interpretation. Humana Press, Totowa, NJ.
16 Borowitz, M. J., Guenther, K. L., Shults, K. E., et al. (1993) Immunophenotyping of acute leukemia by flow cytometric analysis. Use of CD45 and right-angle light scatter to gate on leukemic blasts in three-color analysis. Am. J. Clin. Pathol. 100, 534-540.
17 Dworzak, M. N. and Panzer-Grumayer, E. R. (2003) Flow cytometric detection of minimal residual disease in acute lymphoblastic leukemia. Leuk. Lymphoma 44, 1445-1455.
18 Riley, R. S., Massey, D., Jackson-Cook, C., et al. (2002) Immunophenotypic analysis of acute lymphocytic leukemia. Hematol. Oncol. Clin. North Am. 16, 245-299.
19 Venditti, A., Maurillo, L., Buccisano, F., et al. (2003) Multidimensional flow cyto-metry for detection of minimal residual disease in acute myeloid leukemia. Leuk. Lymphoma 44, 445-450.
Was this article helpful?