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ASCP SCYM study Questions and Answers 100% Solved 2024/2025
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ASCP SCYM study Questions and Answers 100% Solved 2024/2025
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ASCP SCYM studySample Processing - ANSOptimized antibody concentration
Defined range of cell #
Define sample handling, time and temperature
Define acceptable time of delay for acquisition
CD2 - ANST cells: NK cells
CD3 - ANST cells
CD4 - ANST helper cells, monocytes (dimly expressed)
CD5 - ANST cells; Some B Cells
CD8 - ANST cytotoxic cells, macrophages
CD10 - ANSImmature T, B Cells, PMN's
CD13 - ANSMonocytes, PMN's
CD14 - ANSMonocytes, PMN's (dimly expressed)
CD16 - ANSNK cells, PMN's, monocytes
CD19 - ANSB Cells
CD20 - ANSB Cells
CD25 - ANSActivated T Cells
CD33 - ANSMonocytes
CD34 - ANSStem/Progenitor Cells
CD45 - ANSLeukocytes
CD56 - ANSNK cells, monocytes (dimly expressed)
Anti-Kappa - ANSB Cells
Anti-Lambda - ANSB Cells
,Tdt - ANSImmature Lymphocytes
Monoclonal Antibody Staining Direct Staining - ANSRapid single step staining
Can use multiple antibodies from the same host
Monoclonal Antibody Staining Indirect Staining - ANSSecondary amplifies single
A few labeled secondaries can detect many primaries
Clinical Applications of Flow - ANS-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) - ANS2 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 - ANS-Asses degree of immunodeficiency
-Diagnose of AIDS
-Institution of Anti-Retroviral (ARV) therapy and opportunistic infection (OI) prophylaxis
-Monitoring immune reconstitution
CD4 Immunophenotyping Panels - ANS
2 color - ANSIsotype Ctrl
CD45/CD14
CD3/CD4
CD3/CD8
CD3/CD19
CD3/CD16 + 56
3 Color - ANSCD3/CD4/CD45
CD3/CD8/CD45
CD3/CD19/CD16 + 56
4 Color - ANSCD3/CD4/CD8/CD45
CD3/CD16/CD16 + 56/CD45
, Determination of Absolute Counts - ANS-Subset % X Absolute Lymphocyte Count
-Counting Beads- known concentration present in staining tube or added to tube
-Volumetric Cytometers
Leukemia/Lymphoma Phenotyping - ANS-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 - ANS-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 - ANS-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 - ANS-Detection of fetal hemoglobin containing cells in useful for
the diagnosis and management of feto-maternal hemorrhage
-Traditionally assayed with the Kleihauer-Betke method
-FC offers improved sensitivity (<0.02% fetal RBCs) and reproducibility (CV <15%
-Method: Intracellular staining with a fetal hemoglobin specific monoclonal antibody
CD34 Stem Cell Enumeration - ANS-Present on immature hematopoietic cells
-Expression is lost as cells mature
-Used as a marker to identify and enumerate stem cells in bone marrow and peripheral blood
products for transplantation
-Currently the Gold Standard for assessment of HSC products for transplant
CD3 Transplant Monitoring - ANS-Acute graft rejection is an important complication after solid
organ transplantation that may lead to graft loss
-Severe cases of rejection may be treated with antibodies that deplete T cells from the patient
(OKT3, ATG)
-FC is used to assess the effectiveness of T cell depletion after antibody based anti-rejection
therapy
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