ASCP SCYM Study Exam Study Guide.
Sample Processing - answerOptimized antibody concentration
Defined range of cell #
Define sample handling, time and temperature
Define acceptable time of delay for acquisition
CD2 - answerT cells: NK cells
CD3 - answerT cells
CD4 - answerT helper cells, mono...
Sample Processing - answer✔✔Optimized antibody concentration
Defined range of cell #
Define sample handling, time and temperature
Define acceptable time of delay for acquisition
Monoclonal Antibody Staining Direct Staining - answer✔✔Rapid single step staining
Can use multiple antibodies from the same host
Monoclonal Antibody Staining Indirect Staining - answer✔✔Secondary amplifies single
A few labeled secondaries can detect many primaries
Clinical Applications of Flow - answer✔✔-Instrument Performance
-Integrity of the Specimen
-Quality in Processing of Specimens
-Appropriate Reagents and Controls
-Qualified Technical Staff
-Regulatory requirements from CAP, TJC, CLIA, and the FDA exist to ensure the accurate
reporting of flow cytometric results.
Lymphocytes Subset Enumeration (Immunophenotyping) - answer✔✔2 Tubes: 1-
CD3/CD4/CD8/CD45
2- CD3/CD19/CD16&CD56/CD45
-May include analysis of CD4 and CD8 T Cells, B Cells and NK Cells.
-Results reported as proportion (%) of total lymphocyte population.
CD4 Counts in HIV Infection - answer✔✔-Asses degree of immunodeficiency
-Diagnose of AIDS
-Institution of Anti-Retroviral (ARV) therapy and opportunistic infection (OI) prophylaxis
-Monitoring immune reconstitution
CD4 Immunophenotyping Panels - answer✔✔
2 color - answer✔✔Isotype Ctrl
CD45/CD14
CD3/CD4
CD3/CD8
3 Color - answer✔✔CD3/CD4/CD45
CD3/CD8/CD45
CD3/CD19/CD16 + 56
4 Color - answer✔✔CD3/CD4/CD8/CD45
CD3/CD16/CD16 + 56/CD45
Determination of Absolute Counts - answer✔✔-Subset % X Absolute Lymphocyte Count
-Counting Beads- known concentration present in staining tube or added to tube
-Volumetric Cytometers
Leukemia/Lymphoma Phenotyping - answer✔✔-Important in Diagnosis, Classification and
Monitoring of patients with hematologic malignancies.
-Utility based on the fact that hematopoiesis is characterized by the gain and loss of cell surface
and intracellular antigen during the process of maturation.
-Malignant cells may have a block at an immature stage resulting in abnormal numbers of
immune cells. With a mature phenotype may be present in abnormally high numbers. May have
an aberrant phenotype.
Leukemia/Lymphoma Phenotyping FC - answer✔✔-Analysis with panels of monoclonal
antibodies allows for lineage assignment and sub classification that may have further prognostic
significance.
-The identification of a unique phenotype can also facilitate post-therapy monitoring to assess
residual disease.
Relevance for the Diagnosis and Classification of Hematologic Malignancies - answer✔✔-
Expression of aberrant lineage associated antigen
-Absence of lineage associated antigen
-Asynchronous expression of antigens
-Abnormal level of expression of antigen
Detection of Fetal Hemoglobin - answer✔✔-Detection of fetal hemoglobin containing cells in
useful for the diagnosis and management of feto-maternal hemorrhage
-Traditionally assayed with the Kleihauer-Betke method
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