LABS FOR LIFE PROJECT - INDIA

Equipment
Management

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Complete Blood Count Analyzers

These analyzers are used to

count the number of different
types of red and white blood
cells, blood platelets,
hemoglobin, and hematocrit
levels in a blood sample.

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Types of Cell Counters

3-Part Differential Cell Counters

– Provides neutrophils, lymphocytes
and mononuclear cell counts Erythrocytes Neutrophils
– Unable to distinguish between
monocyte, eosinophils, and basophils.
5-Part Differential Cell Counters Lymphocytes Basophils

– Able to determine granularity,

diameter, and inner complexity of Monocytes Eosinophils

blood cells.
– Able to differentiate all 5 population Platelets
of WBCs
– Some cell counters are able to detect
and abnormal cell populations

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Principle of operation of Cell counter

Impedance Principle:
• Cells are non-conductive
• Electrical resistance caused by
cells is proportional to their
volume /size.
• Various blood cells could be
differentiated based on this
principle

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Principle of operation of Cell counter

• Diluted blood is passed between

two electrodes through an aperture
• Impedance changes as a cell passes
through is monitored.
• A lytic reagent is added to the blood
solution to selectively lyse the red
cells (RBCs) leaving only white cells
(WBCs), and platelets intact.
• The platelet count is easily
separated from the WBC count by
the smaller impedance spikes they
produce in the detector due to their
lower cell volumes
• Impedance principle is used for
CBCs and three-part WBC
differentials

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 The Impedance principle

Red Blood Cell A red cell passes

through RBC aperture

Sensing Zone

Oscilloscope
Oscilloscope
 The Impedance principle

Red Blood Cell A red cell passes

through RBC aperture

Sensing Zone

Oscilloscope
Oscilloscope
 The Impedance principle

Neutrophil A White cell passes

through WBC aperture

Oscilloscope

Sensing Zone

Oscilloscope
 The Impedance principle

Neutrophil A White cell passes

through WBC aperture

Oscilloscope

Sensing Zone

Oscilloscope
 The Impedance principle

Neutrophil A White cell passes

through WBC aperture

Oscilloscope

Sensing Zone

Oscilloscope
Applying Thresholds to separate Plt from RBC

20 fl

2 fl

Base line

Platelets
RBC of various
sizes
 3-Part Differential Histogrm

Lymphocyte Monocyte Granulocyte

256 channel high resolution WBC histogram

display cell population data between 35 and 450 fl
Coulter WBC histogram

Monocytes
Lymphocytes

Neutrophils

Basophils Eosinophils
Principle of operation of 5-part Differential Counters

• Besides using impedance principle, 5-part

differential cell counters employ
flowcytometric principles
• Through hydrodynamic focusing cells are
passed through an aperture in single file
• Cells are subjected to laser beam which
assists in:
– Identifying granularity of the cells
– Identify nuclear structure
• Based on size, granularity and nuclear
structure, cells are separated in 3-
dimentional space
• All five types of WBCs are identified based
on their typical position in 3-dimentional
space

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Hemoglobin Measurement

WBC Bath Signal

ADC
Processor
HGB Lamp

Derivation
Absorbance= log 10 ( VR / Vs )

Where VR = Reference voltage

Vs = Sample voltage

HGB Sensor &

Sample PRE-AMP

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Cell counter – Key components

Tube cap piercer

Typical report
Single Sample port

Tube rack station

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Maintenance of Cell Counters
Follow your manufacturer’s instructions manual for specific maintenance needs of your
analyzer – instructions given here are only indicative
• Always use PPE while performing maintenance
• Keep the cell counter surface and surroundings clean using 10% bleach.
• Most of computerized cell counters have computer-controlled maintenance menu.
Users must be familiar with these
• Inspect the bottom of washing head for salt build up – should be wiped off with a
damp cloth or wiper
• Check tubing system and look for any liquid leakage. If you experience leakage,
contact authorized technician.
• Perform daily start-up and shut-down cycles – these cycles are important for auto-
cleaning of tubing and apertures and hence in reducing blank values
• Check reagent status and change if required
• Empty waste container before starting the day
Waste contains human origin substances representing biohazard.
Neutralization of biohazard waste:
• Put 2 ml per liter of hypochlorite solution into the waste. Close the cap and
• Shake the container., after 1 hour you can dispose of the Waste liquid into the drain.

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 Example of Weekly Maintenance of Cell Counters
Follow your manufacturer’s instructions manual for specific maintenance needs of your
analyzer – instructions given here are only indicative

• Cleaning of Probe washing head:

• Salt build-up on the lower surface of wash
head may cause malfunction during
operation leading to carryover between
samples.
• Use a soft cloth or wiper dampened with Probe wash head
water to clean this area.
• Inspect aspiration / dilution syringes for leaks or
salt buildup
• Clean sample auto-loader station and track

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Calibration of cell counters
Automated hematology analyzers shall be calibrated using calibrator provided by
manufacturer: Controls shall not be used for calibration.
• Hematology calibrators have short expiry and very short open vial
stability.
• Ensure traceability of calibrators to ICSH.
• Some 3 part equipment calibrators have a defined Uncertainty of
Measurement and give a range for the target.
• Calibrators with more accurate values give a single, equipment
specific target value.
• It is important to ensure adequate equipment performance in
terms of precision, background values and carry over values
before performing CBC analyzer calibrations.
• Both closed and open modes should be calibrated whenever 2
modes are available.
• Calibration should be followed by verification QC to verify
acceptable performance
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