Identification of Leukocyte Populations Using EPICS XL Flow Cytometer and Microscopy
Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
Leukemia is an uncontrolled growth of abnormal white blood cells both in blood and bone marrow. Depending on the type of leukocytes that dominates, it can be divided in lymphoid and myeloid leukemia. Diagnosis is performed by differential counting of leucocytes in a blood smear utilizing microscopy or immunophenotypning and flow cytometry. Identification by microscopy is time consuming and requires much experience. It is also impossible to distinguish between B- and T-lymphocytes, which flow cytometry can do. With flow cytometry it is possible to analyze thousands of cells in a short time. The goal of this work was to identify different leukocyte populations in blood from patients with hematologic malignancies. The study used an EPICS XL flow cytometer with four fluorescence channels, which can analyze seven different antibodies with four different fluorochromes simultaneously. Patient samples were analyzed and each cell type in the blood smears was compared with one or more corresponding CD markers in flow cytometry. The stability of prepared samples was examined by analyzing each sample twice within 24 hours. The study also analyzed cells that were isolated by Ficoll separation. The study showed that there is a good correlation between proportion of lymphocytes in blood smears and the proportion of cells expressing the CD2 and CD19 in flow cytometry. However, it was difficult to find a direct link between the immature stages of leukocytes in blood smears and various CD markers in flow cytometry. The sample was stable for 24 hours at +4 C.
Place, publisher, year, edition, pages
Flow cytometry, leukocyte differential, Fluorochrome, leukemia, CD markers
IdentifiersURN: urn:nbn:se:uu:diva-154795OAI: oai:DiVA.org:uu-154795DiVA: diva2:422311