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Automated Blood cell analyzers
the basics
JG Nel
Department of Haematology
• (picture of the road ahead)
• (rearview mirror)
• (microscope)
Wallace and Joseph Coulter
• (Wallace and Joseph Coulter)
The Coulter model A
The Red cell count
• (red cells)
Aperture impedance counting
Light scattering
Factors influencing the reliability of
automated counts
• Standardisation
• Coincidence counting
Coincidence counting
Hydrodynamic focussing
Correcting for coincidence
Factors influencing the reliability of
automated counts
• Standardisation
• Coincidence counting
• Diluents
• Discrimination thresholds
• The counting technique employed
The MCV and Haematocrit
• MCV – can be: derived directly
or derived from Hct and
RCC
• Haematocrit derived directly
or derived from MCV
and RCC
MCH and MCHC
• MCH : a derived parameter, using
Hb and RBC
• MCHC: derived from the Hb,Hct and
sometimes the MCV
Red cell Distribution Width
• Equivalent to the microscopic assessment
of the degree of anisocytosis
• Of some value in distinguishing
1.Fe deficiency anaemia
Thallasaemia
2.Megaloblastic anaemia
Other causes Macrocytosis
The WCC
• Determined by Impedance counting or
Light scattering
• Thresholds set to exclude normal platelets
• Nucleated cells usually included
Sources of factitiously low WCC
• Leukocyte agglutination
• Prolonged sample storage
• Abnormally fragile cells
Sources of factitiously high WCC
• Usually the result of failure of red cells to
lyse
• Micro clots
• Platelet clumping
• The presence of cryoglobulin
Automated differential counts
• Three principles used in various systems
1. Computerized pattern recognition
2. Flow cytochemistry
3. Leukocyte volume analysis
Computerized pattern recognition
• (computer)
Flow cytochemistry
Leukocyte volume analysis
Counting platelets
Sources of factitious platelet counts
• High counts: markedly microcytic red cells
fragmented red cells
• Low counts: Giant platelets
EDTA induced platelet
clumping
Satelitism
Counting reticulocytes
Counting Retics
The Hb
• Red cells lysed
• Free Hb converted to
haemoglobinsulphate
• Spectrophotometry
Quality control
• Accuracy
• Precision
The place of the automated blood
cell analyzer
• Cannot recognize all the abnormalities
that a human observer can
• Designed to produce accurate and precise
counts
• Alert the operator when a specimen has
unusual characteristics
Flagging
• Results should be flagged when:
1. The specimen might contain: blasts,
immature granulocytes, NRBC or
atypical Lymphocytes
2. Giant or aggregated platelets are present
or distinction between the RBC and Plt
populations cannot be made
3. Abnormalities present assoc with
factitious results
A result is can never be better than
the specimen
Automated blood cell analisers

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Automated blood cell analisers