Med
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Hematology and Health Technology Research Center Faculty of Associated Medical Sciences Chiang Mai University
Automated CBC • RBC, MCV counted automatically, but Hct derived • Hb determined automatically, but MCH and MCHC derived • Platelet counted automatically • WBC differential count : 3 part (Lymp, Mid, Gran) or 5 part (Lymp, PMN, Mono, Eo, Baso) • Blood film : RBC morpho and to confirm WBC diff and plt count • RDW : Degree of anisocytosis
Electrical-impedance method of cell counting
Light-scatter method of cell counting
Histogram of three-part differential count
Scattergram of five-part differential count
Hematology analyzers Techniques Brands Elec-impedance Coulter ONYX Coulter AC-T diff Serono-Baker Light-scatter Coulter VCS, STKS, GEN-S Bayer ADVIA-120 Combination ABBOTT Cell-Dyn 3500
Diff. 5-part 3-part 3-part 5-part 5-part 5-part
Flow Cytometry • Measurement of properties • Single particles or cells • Suspend in fluid • Flow singly through a beam of light
Cellular Properties • Direct –Size –Granularity • Indirect –Fluorescence Stain –Immunofluorescent Stain
Forward Scatter (Size)
Side Scatter (Granules)
Flow Cytometer • Fluidic System • Optical System • Detector System • Data Processing System
4 Principle & compartments of Flow Cytometer
1 2 3
Flow Cytometry for WBCs • Immunophenotyping of Lymphocytes • Leukemia Diagnosis (typing, cell cycle, ploidy status etc.) • Minimal Residual Disease (MRD)Detection
Immunophenotyping • Surface markers (antigens) – CDs – sIg (B-cells) • Cytoplasmic makers – cIg (preB-cells) – TdT (early lymphoid cells) • Flow Cytometry vs Fluorescent Microscopy
Comparison of Flow Cytometry and Fluorescent Microscopy
Automated CBC • RBC, MCV counted automatically, but Hct derived • Hb determined automatically, but MCH and MCHC derived • Platelet counted automatically • WBC differential count : 3 part (Lymp, Mid, Gran) or 5 part (Lymp, PMN, Mono, Eo, Baso) • Blood film : RBC morpho and to confirm WBC diff and plt count • RDW : Degree of anisocytosis
Electrical-impedance method of cell counting
Light-scatter method of cell counting
Histogram of three-part differential count
Scattergram of five-part differential count
Hematology analyzers Techniques Brands Elec-impedance Coulter ONYX Coulter AC-T diff Serono-Baker Light-scatter Coulter VCS, STKS, GEN-S Bayer ADVIA-120 Combination ABBOTT Cell-Dyn 3500
Diff. 5-part 3-part 3-part 5-part 5-part 5-part
Flow Cytometry • Measurement of properties • Single particles or cells • Suspend in fluid • Flow singly through a beam of light
Cellular Properties • Direct –Size –Granularity • Indirect –Fluorescence Stain –Immunofluorescent Stain
Forward Scatter (Size)
Side Scatter (Granules)
Flow Cytometer • Fluidic System • Optical System • Detector System • Data Processing System
4 Principle & compartments of Flow Cytometer
1 2 3
Flow Cytometry for WBCs • Immunophenotyping of Lymphocytes • Leukemia Diagnosis (typing, cell cycle, ploidy status etc.) • Minimal Residual Disease (MRD)Detection
Immunophenotyping • Surface markers (antigens) – CDs – sIg (B-cells) • Cytoplasmic makers – cIg (preB-cells) – TdT (early lymphoid cells) • Flow Cytometry vs Fluorescent Microscopy
Comparison of Flow Cytometry and Fluorescent Microscopy
