Siemens (formerly Bayer) Advia 120. 120: CBC and Retic: ▫WBC, RBC and PLT
counts are obtained using optical scatter. ▫Performs WBC count by two methods.
Hematology Analyzers Automated hematology analyzers are complex and incorporate multiple technologies within one instrument.
CLS417 - CLINICAL HEMATOLOGY II Linda Sykora, MT(ASCP)SH
This advanced technology requires that the clinical laboratory scientist possess the interpretive skills to evaluate computer-analyzed data, including cell distribution plots, and have a good understanding of instrument limitations.
Rotation II Auto Unit
Automated Cell Counting and Differential Systems Copyright © University of Nebraska 2008 1
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A brief overview of current analyzers will be presented. During the presentation,
Hematology Analyzers
All multimulti-parameter cell counting systems:
The implementation of accurate automated differentials has been a complicated process. Flow cytometry has evolved as the primary technology for automated WBC differentials.
▪Utilize the electronic impedance and/or optical scatter principle of operation ▪Measure hemoglobin photometrically ▪Count red cells and platelets together ▪Employ hydrodynamic focusing ▪Generally produce reliable CBC results
In some instruments, the optical flow cell is also incorporated into CBC measurements. Often, more than one method is used in reporting a single parameter, i.e., duplicate WBC counts or PLT counts are done, which increases reliability when interference exists. 4
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True or False. The main WBC differential plot generated by all analyzers shows a similar cell distribution pattern. pattern
Advances in the latest instruments include: ▪Addition of stains, dyes or fluorescence
measurement ▪Enumeration of nucleated red cells ▪Expanded parameter linearity limits ▪Automated Retic counts and immature reticulocyte fraction (IRF) ▪Dual counts for certain parameters ▪Sophisticated instrument and sample flags
Absorption
LH750
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CLS417 Clinical Hematology II Rot II Auto Unit Automated Cell Counting and Differential Systems
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True. The location of normal cell types is comparable regardless of the differential technology utilized.
Siemens (formerly Bayer) Advia 120: 120 CBC and Retic: ▪WBC, RBC and PLT counts are obtained using optical scatter ▪Performs WBC count by two methods ▪Uses nucleic acid dye for retic counting
Neuts Monos
Monos
Neuts
Lymphs
Lymphs
Eos Eos Absorption
Differential (Flow cytometry, Cytochemistry): ▪Stains for peroxidase and measures light scatter and absorption to identify WBC types ▪Strips the cytoplasm of all cells and uses light scatter to identify basophils 8
LH750 Monos
Neuts
Monos
Lymphs Lymphs
Neuts
Eos Eos
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Normal WBC cytogram distribution
Bayer Advia 120 Normal Printout
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‘Messy’ neutrophil cluster
Bayer Advia 120 Abnormal Printout Neutrophilia with left shift and immature gran (IG) flags. Needs manual differential.
Normal CBC & Diff parameters
No flags
Normal RBC/PLT histograms 9
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Normal WBC scatterplot distribution
Abbott CellCell-Dyn 4000 & Sapphire: CBC and Retic: ▪WBC, RBC and PLT counts are obtained using impedance and/or light scatter ▪Performs RBC and PLT by two methods ▪Uses RNA fluorescence for retic counting
Normal CBC & Diff parameters
Note two RBC & PLT counts; can quantitate NRBCs
Differential (MAPSS technology): ▪Measures 4 angles of light scatter…0o, 7o, 90o, 90o depolarized…to identify WBC types ▪Enumerates nucleated rbcs using DNA fluorescence 11
Normal RBC/PLT histograms & scatterplot 12
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CLS417 Clinical Hematology II Rot II Auto Unit Automated Cell Counting and Differential Systems
No flags
Cell-Dyn 4000 Normal Printout
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Beckman Coulter LH series: Cell-Dyn 4000 Abnormal Printout Macrocytosis and NucRBCs. NucRBCs Note abnormal WBC scatterplot pattern. Needs manual differential.
CBC and Retic: ▪WBC, RBC and PLT counts are obtained using impedance ▪Uses new methylene blue for retic counting
NRBCs
RBC histogram is shifted to the right.
NRBCs
Differential (VCS technology): ▪Simultaneously measures cell volume, conductivity and light scatter to identify WBC types ▪Enumerates nucleated rbcs using cell cluster analysis
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Coulter LH500 Abnormal Printout
Coulter LH750 Normal Printout
Critical high WBC & critical low PLT. All diff parameters are flagged.
Normal CBC & Diff parameters Can quantitate NRBCs Normal RBC/PLT/WBC histograms
Blasts (pattern ‘streams’ upward due to large cells)
Blast flags and WBC scatterplot pattern suggests acute leukemia. leukemia Needs manual differential and PLT estimate.
No flags
Normal WBC scatterplot distribution 15
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Sysmex XE series:
Normal WBC scatterplot distribution
Sysmex XE Normal Printout
CBC and Retic: ▪WBC, RBC and PLT counts are obtained using direct current (impedance) and/or light scatter ▪Performs WBC and PLT by two methods ▪Uses fluorescent dye for retic counting
No flags
Note two PLT methods; can quantitate NRBCs Normal CBC & Diff parameters
Differential (Flow Cytometry, Fluorescence): ▪Uses light scatter, fluorescent light and RF/DC technology to identify WBC types ▪Enumerates nucrbcs using fluorescence 17
Normal RBC/PLT histograms 18
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CLS417 Clinical Hematology II Rot II Auto Unit Automated Cell Counting and Differential Systems
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Evaluate the patient printout. What do you suspect?
Sysmex XE Abnormal Printout Flagged WBC, RBC, and PLT data. All diff parameters are flagged and NRBCs are present. The WBC is corrected for nucRBCs. Needs manual differential.
NRBCs
The optical PLT result is on the printout; the impedance PLT
histogram shows interference. A severe microcytic anemia is present as well as marked anisocytosis (high RDW).
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Evaluate the patient printout. What is present at the arrow?
Chronic lymphocytic leukemia Critical high WBC, normal Hgb & PLT, relative & absolute lymphocytosis.
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Lymphs
Smudge cells
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Nucleated red cells
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SUMMARY:
Uncorrected WBC of 29.3 K/uL (includes NRBCs)
CBC and Retic Parameters ▪Siemens (formerly Bayer) Advia 120 WBC, RBC and PLT counts are obtained using light scatter; uses nucleic acid dye for retic counting
WBC of 23.7K/uL does not include NRBCs (corrected).
▪Abbott Cell-Dyn 4000 & Sapphire WBC, RBC and PLT counts are obtained using impedance and/or light scatter; RNA fluorescence is used for retic counts ▪Beckman Coulter LH series WBC, RBC and PLT counts are obtained using impedance; uses new methylene blue for retic counting
NRBCs
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CLS417 Clinical Hematology II Rot II Auto Unit Automated Cell Counting and Differential Systems
▪Sysmex XE series WBC, RBC and PLT counts are obtained using direct current (impedance) and/or light scatter; uses fluorescent dye for retic 24 counts
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SUMMARY: Differential Technology ▪Siemens Advia 120 – Flow Cytometry & Cytochemistry Stains for peroxidase and measures light scatter and absorption to identify WBC types
▪Cell-Dyn 4000/Sapphire – MAPSS Measures 4 angles of light scatter to identify WBC types
▪Beckman Coulter LH – VCS Simultaneously measures cell volume, conductivity and light scatter to identify WBC types
▪Sysmex XE –Flow Cytometry, Fluorescence & RF/DC Measures light scatter, fluorescence, and RF/DC to identify WBC types 25
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CLS417 Clinical Hematology II Rot II Auto Unit Automated Cell Counting and Differential Systems
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