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    Automated Cell Counting Instrumentation

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    Cecille Ann
    Automated cell counters use various principles like impedance, light scattering, fluorescence to provide a complete blood count and differential within minutes. Parameters are directly measured or calculated based on other parameters. Different instruments may use similar principles but implement them differently. Histograms and cytograms can identify abnormalities and sources of error for corrective action.

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    Automated Cell Counting Instrumentation

    Uploaded by

    Cecille Ann
    255 views33 pages
    Automated cell counters use various principles like impedance, light scattering, fluorescence to provide a complete blood count and differential within minutes. Parameters are directly measured or calculated based on other parameters. Different instruments may use similar principles but implement them differently. Histograms and cytograms can identify abnormalities and sources of error for corrective action.

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    Automated Cell Counting Instrumentation (1)

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    Automated cell counters use various principles like impedance, light scattering, fluorescence to provide a complete blood count and differential within minutes. Parameters are directly measured or calculated based on other parameters. Different instruments may use similar principles but implement them differently. Histograms and cytograms can identify abnormalities and sources of error for corrective action.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    255 views33 pages

    Automated Cell Counting Instrumentation

    Uploaded by

    Cecille Ann
    Automated cell counters use various principles like impedance, light scattering, fluorescence to provide a complete blood count and differential within minutes. Parameters are directly measured or calculated based on other parameters. Different instruments may use similar principles but implement them differently. Histograms and cytograms can identify abnormalities and sources of error for corrective action.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 33

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    Explain the different principles of automated cell counting. Describe how the general principles are implemented on the different instruments discussed. Identify the hemogram parameters directly measured by analyzers. Explain the derivation of calculated or indirectly measured hemogram parameters. Explain the derivation of the WBC differential count. Interpret and compare patient data, including WBC and RBC histograms or cytograms or both, obtained from the major hematology instruments. Explain the general principles of automated reticulocyte counting. Identify sources of error in automated cell counting and determine appropriate corrective action.

    AUTOMATION provides greater accuracy and

    precision than manual methods.


    Hematology analyzers provide the eight standard

    hematology parameters (CBC) plus a three-part or five-part differential leukocyte count in less than 1 minute on 200 uL of whole blood.
    Automation allows more efficient workload

    management and more timely diagnosis and treatment of disease.

    Low voltage direct current (DC) resistance Developed by Coulter in the 1950s PRINCIPLE : Cell counting is based on the detection

    and measurement of changes in electrical resistance produced by cells as they traverse a small aperture.

    Aperture Diameter
    RBC/Platelet aperture is smaller than the WBC aperture

    to increase platelet counting sensitivity Protein build up : decreases the diameter of the orifice, slowing the flow of cells
    Burn Circuits or Internal Cleaning Systems
    Minimize protein build up and carry over of cells from

    one sample to the next


    Coincident Passage
    Causes count reduction

    Orientation of the cell in the center of the aperture

    and deformability of the RBC


    Recirculation of cells back in the sensing zone creates

    erroneous pulses and falsely elevated cell counts


    Hydrodynamic focusing
    Avoids many potential problems

    Alternating current resistance Low voltage DC maybe used in conjunction with RF

    resistance, or resistance to a high voltage electromagnetic current flowing between both electrodes simultaneously
    The total volume of the cell is proportional to the

    change in DC, the cell interior density is proportional to pulse size or change in the RF signal.

    CONDUCTIVITY : attenuated by nucleus to

    cytoplasm ratio, nuclear density, and cytoplasmic granulation

    Two dimensional distribution cytogram Created by plotting impedance against conductivity Displays cell populations as clusters, with the number

    of dots in each cluster representing the concentration of that cell type.

    Uses both laser and non-laser light Latest technology

    Hydraulics System
    Includes aspirating unit, dispensers, diluters, mixing

    chambers, aperture baths, or flow cells or both, and a hemoglobinometer


    Pneumatics System
    Includes vacuums and pressures required for operating

    the valves and moving the sample through the hydraulics system
    Electrical Systems
    Controls operational sequences of the total system and

    includes electronic analyzers and computing circuitry for processing the data gathered

    Parameter

    Principle

    WBC
    RBC Hb Hct MCV

    Impedance, hydrodynamic focusing


    Impedance Modified Cyanmethemoglobin at 525 nm (RBC x MCV)/10 Mean of RBC volume histogram

    MCH
    MCHC

    (Hb/RBC) X 10
    (Hb/Hct) X 100

    Platelet Count Impedance (2-20 fL) : least squares fit of volume distribution histogram (0-70 fL)
    RDW CV (%) of RBC histogram : (SD/MCV) X 100

    Reticulocyte Count

    NMB-N, volume, conductivity, optical scatter

    Parameter

    Principle

    WBC
    RBC Hb Hct MCV

    Hydrodynamic Focusing, DC Detection (impedance)


    Hydrodynamic Focusing, DC Detection (impedance) SLS Hb (555 nm) Cumulative Pulse Height Detection (Hct/RBC) X 10

    MCH
    MCHC

    (Hb/RBC) X 10
    (Hb/Hct) X 100

    Platelet Count
    RDW

    Hydrodynamic focusing, DC detection (impedance) approximately 2 30 fL


    RDW-SD (fL) or RDW-CV (%) available

    Reticulocyte Count

    Auramine O; fluorescence detection

    Parameter

    Principle

    WBC
    RBC Hb Hct MCV

    Optical Scatter (primary count); Impedance (secondary count)


    Impedance Modified Cyanmethemoglobin (540 nm) (RBC X MCV)/ 10 Mean of RBC distribution histogram

    MCH
    MCHC

    (Hb/RBC) X 10
    (Hb/Hct) X 10

    Platelet Count
    RDW

    Impedance (approximately 2 30 fL)


    Relative value, equivalent to CV

    Reticulocyte Count

    Propriety Stain (CD4K530), multiangular scatter, and fluorescence detection

    Parameter

    Principles

    WBC
    RBC

    Hydrodynamic focusing, optical scatter and absorption


    Hydrodynamic focusing, laser low-angle (2 3 degree) and highangle (5-15 degree) scatter

    Hb
    Hct

    Modified Cyanmethemoglobin (546 nm)


    (RBC X MCV)/10

    MCV
    MCH MCHC Platelet Count

    Mean of RBC volume histogram


    (Hb / RBC) X 10 (Hb/Hct) X 100 Hydrodynamic focusing, laser low-angle (2 3 degree) and highangle (5-15 degree) scatter (1-60 fL)

    RDW
    Reticulocyte Count

    CV (%) of RBC histogram: (SD/MCV) X 100


    Oxazine 750; low-angle (2-3 degree) and high-angle (5-15 degree) opticla scatter and absorbance

    Parameter Neutrophil

    Coulter VCS

    Sysmex OS and FS

    Abbott MAPSS

    Siemens PS, OS, Abs

    Lymphocyte Monocytes
    Eosinophils Basophils

    VCS VCS
    VCS VCS

    OS and FS OS and FS
    OS and FS OS and FS

    MAPSS MAPSS
    MAPSS MAPSS

    PS, OS, Abs PS, OS, Abs


    PS, OS, Abs Differential lysis, laser low-angle (2-3 degree) and high angle (5-15 degree) scatter

    Parameters Affected

    RBC decreased, MCV, MCHC increased, grainy appearance

    Rationale Instrument Indicators


    Corrective Action

    Agglutination of RBCs Dual RBC population on RBC map, or right shift on RBC histogram
    Warm sample to 37 degrees C and rerun

    Parameters Affected

    Hb, MCH increased

    Rationale

    Increased turbidity affects spectrophotometric readings


    Hb X 3 is not Hct+/-3, abnormal histogram

    Instrument Indicators

    Corrective Action

    Plasma replacement

    Parameters

    RBC, Hct decreased

    Rationale
    Instrument Indicators

    RBCs lysed and can not be counted


    Hb X 3 is not Hct+/-3, may show lipemia pattern on histogram

    Corrective Action

    Request new sample

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