ASCP SCYM study question with answers
Sample Processing - Optimized antibody concentration
Defined range of cell #
Define sample handling, time and temperature
Define acceptable time of delay for acquisition
-CD2 - T cells: NK cells
-CD3 - T cells
-CD4 - T helper cells, monocytes (dimly expressed)
-CD5 - T cells; Some B Cells
-CD8 - T cytotoxic cells, macrophages
-CD10 - Immature T, B Cells, PMN's
-CD13 - Monocytes, PMN's
-CD14 - Monocytes, PMN's (dimly expressed)
-CD16 - NK cells, PMN's, monocytes
-CD19 - B Cells
-CD20 - B Cells
-CD25 - Activated T Cells
-CD33 - Monocytes
-CD34 - Stem/Progenitor Cells
-CD45 - Leukocytes
-CD56 - NK cells, monocytes (dimly expressed)
-Anti-Kappa - B Cells
-Anti-Lambda - B Cells
,-Tdt - Immature Lymphocytes
-Monoclonal Antibody Staining Direct Staining - Rapid single step staining
Can use multiple antibodies from the same host
-Monoclonal Antibody Staining Indirect Staining - Secondary amplifies single
A few labeled secondaries can detect many primaries
-Clinical Applications of Flow - -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) - 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 - -Asses degree of immunodeficiency
-Diagnose of AIDS
-Institution of Anti-Retroviral (ARV) therapy and opportunistic infection (OI) prophylaxis
-Monitoring immune reconstitution
-CD4 Immunophenotyping Panels -
-2 color - Isotype Ctrl
CD45/CD14
CD3/CD4
CD3/CD8
CD3/CD19
CD3/CD16 + 56
-3 Color - CD3/CD4/CD45
CD3/CD8/CD45
CD3/CD19/CD16 + 56
-4 Color - CD3/CD4/CD8/CD45
CD3/CD16/CD16 + 56/CD45
-Determination of Absolute Counts - -Subset % X Absolute Lymphocyte Count
-Counting Beads- known concentration present in staining tube or added to tube
, -Volumetric Cytometers
-Leukemia/Lymphoma Phenotyping - -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 - -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 - -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 - -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 - -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 - -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
-Organ Transplantation Antibody Detection - -The presence of HLA specific antibodies in the
potential recipient of a kidney transplant that are specific for the potential donor is often a
contraindication to transplantation.
Sample Processing - Optimized antibody concentration
Defined range of cell #
Define sample handling, time and temperature
Define acceptable time of delay for acquisition
-CD2 - T cells: NK cells
-CD3 - T cells
-CD4 - T helper cells, monocytes (dimly expressed)
-CD5 - T cells; Some B Cells
-CD8 - T cytotoxic cells, macrophages
-CD10 - Immature T, B Cells, PMN's
-CD13 - Monocytes, PMN's
-CD14 - Monocytes, PMN's (dimly expressed)
-CD16 - NK cells, PMN's, monocytes
-CD19 - B Cells
-CD20 - B Cells
-CD25 - Activated T Cells
-CD33 - Monocytes
-CD34 - Stem/Progenitor Cells
-CD45 - Leukocytes
-CD56 - NK cells, monocytes (dimly expressed)
-Anti-Kappa - B Cells
-Anti-Lambda - B Cells
,-Tdt - Immature Lymphocytes
-Monoclonal Antibody Staining Direct Staining - Rapid single step staining
Can use multiple antibodies from the same host
-Monoclonal Antibody Staining Indirect Staining - Secondary amplifies single
A few labeled secondaries can detect many primaries
-Clinical Applications of Flow - -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) - 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 - -Asses degree of immunodeficiency
-Diagnose of AIDS
-Institution of Anti-Retroviral (ARV) therapy and opportunistic infection (OI) prophylaxis
-Monitoring immune reconstitution
-CD4 Immunophenotyping Panels -
-2 color - Isotype Ctrl
CD45/CD14
CD3/CD4
CD3/CD8
CD3/CD19
CD3/CD16 + 56
-3 Color - CD3/CD4/CD45
CD3/CD8/CD45
CD3/CD19/CD16 + 56
-4 Color - CD3/CD4/CD8/CD45
CD3/CD16/CD16 + 56/CD45
-Determination of Absolute Counts - -Subset % X Absolute Lymphocyte Count
-Counting Beads- known concentration present in staining tube or added to tube
, -Volumetric Cytometers
-Leukemia/Lymphoma Phenotyping - -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 - -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 - -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 - -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 - -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 - -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
-Organ Transplantation Antibody Detection - -The presence of HLA specific antibodies in the
potential recipient of a kidney transplant that are specific for the potential donor is often a
contraindication to transplantation.
