Sample Processing - ANSWER Optimized antibody concentration
Defined range of cell #
Define sample handling, time and temperature
Define acceptable time of delay for acquisition
CD2 - ANSWER T cells: NK cells
CD3 - ANSWER T cells
CD4 - ANSWER T helper cells, monocytes (dimly expressed)
CD5 - ANSWER T cells; Some B Cells
CD8 - ANSWER T cytotoxic cells, macrophages
CD10 - ANSWER Immature T, B Cells, PMN's
CD13 - ANSWER Monocytes, PMN's
CD14 - ANSWER Monocytes, PMN's (dimly expressed)
CD16 - ANSWER NK cells, PMN's, monocytes
1
,CD19 - ANSWER B Cells
CD20 - ANSWER B Cells
CD25 - ANSWER Activated T Cells
CD33 - ANSWER Monocytes
CD34 - ANSWER Stem/Progenitor Cells
CD45 - ANSWER Leukocytes
CD56 - ANSWER NK cells, monocytes (dimly expressed)
Anti-Kappa - ANSWER B Cells
Anti-Lambda - ANSWER B Cells
Tdt - ANSWER Immature Lymphocytes
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
2
, 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.
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 re-
porting of flow cytometric results.
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
CD3/CD19
CD3/CD16 + 56
3
Defined range of cell #
Define sample handling, time and temperature
Define acceptable time of delay for acquisition
CD2 - ANSWER T cells: NK cells
CD3 - ANSWER T cells
CD4 - ANSWER T helper cells, monocytes (dimly expressed)
CD5 - ANSWER T cells; Some B Cells
CD8 - ANSWER T cytotoxic cells, macrophages
CD10 - ANSWER Immature T, B Cells, PMN's
CD13 - ANSWER Monocytes, PMN's
CD14 - ANSWER Monocytes, PMN's (dimly expressed)
CD16 - ANSWER NK cells, PMN's, monocytes
1
,CD19 - ANSWER B Cells
CD20 - ANSWER B Cells
CD25 - ANSWER Activated T Cells
CD33 - ANSWER Monocytes
CD34 - ANSWER Stem/Progenitor Cells
CD45 - ANSWER Leukocytes
CD56 - ANSWER NK cells, monocytes (dimly expressed)
Anti-Kappa - ANSWER B Cells
Anti-Lambda - ANSWER B Cells
Tdt - ANSWER Immature Lymphocytes
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
2
, 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.
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 re-
porting of flow cytometric results.
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
CD3/CD19
CD3/CD16 + 56
3
