Electrical Impedance and Colorimetric Measurements

Electrical Impedance
and
Colorimetric Measurements
Joanna Ellis,
MLS(ASCP)
Objectives
Cite the electrical impedance
principle of cell counting.
Identify and interpret microcytic and
macrocytic RBC histograms
Define coincidence
Identify the cell populations
represented on a 3-part differential
WBC histogram
History
You CAN patent a hole
Prior to the 1950s blood cell counts were
performed by manual methods:
Hemacytometer blood counts

Spun hematocrits
Spectrophotometrically determined hemoglobins
Peripheral blood cell evaluation for all differentials
In 1953, Wallace Coulter patented the Coulter

Principle in which particles are counted in fluid
passed through a hole. The incredulous
attorneys who had told him You cant patent a
hole were proven wrong.
Hematology automation has since grown to
include additional principles such as optical
light scatter and flow cytometry.
The Coulter Principle

The poorly conductive blood cells are suspended in
a conductive diluent (liquid).
The diluent is passed through an electric field
created between two electrodes.
The liquid passes through a small aperture (hole).
The passage of each particle through the aperture
momentarily increases the impedance (resistance)
of the electrical path between the electrodes.
The increase in impedance creates a pulse that
can be measured.
The number of pulses = blood cell count
The amplitude (height) of the pulse = Volume of
cell
Electrical Impedance (The Coulter

Principle)
Sweep Flow: Steady stream of diluent that flows behind each

aperture to prevent cells from re-entering the aperture
Counting Chambers
RBC/Platelet
Chamber
WBC Chamber
Differential
Chamber
Reticulocyte
Channel
Most common chambers using impedance:

RBC/Platelet chamber
WBC chamber
RBC/Platelet Chamber
Particle Size
Cell the instrument

Counts
2-20 femtoliters (fL)
Platelet
36-360 fL
Red Blood Cell
RBC and Platelet

Histograms
The black line represents normal cell distribution. The

red line on the RBC histogram graphically represents
a microcytic red cell population.
Bimodal Histogram
Bimodal peak can be seen in situations such
Cold agglutinin disease
Post-transfusion
Post-treatment of IDA
Coincidence
Coincidence: Multiple cells passed through

the
aperture at once.
WBC Chamber
WBCs
Lysing
agent
Hgb
released
Aperture in
electric
field
In some analyzers the WBC count is directly

measured by electric impedance after the red cells
have been destroyed by a lysing agent. The lysing
agent also shrinks the leukocyte cell membrane
and cytoplasm; therefore, the WBC count
represents the measure of the cell volume not
Colorimetric
Measurements
Hemoglobin is often determined by a colorimetric method.
Imidazole
Non-cyanide reagent with color change and read at
540nm
Instruments
Abbott CELL-DYN Sapphire
Sodium Lauryl Sulfate
Non-cyanide reagent with color change and read
spectrophotometrically
Instruments
Sysmex XT and XE
Lysing agent converts free hemoglobin to
cyanmethemoglobin and read spectrophotometrically at
540nm.
Instruments
Advia 120
Some Beckman Coulters
Three-part Differential
WBC Histogram
Lymphocytes: 35-92L
Mononuclear Cells: 92-152 fL
Granulocytes: 152-450 fL
References
"Abaxis Veterinary Diagnostics - VetScan HM2 Technology." Abaxis Medical

Diagnostics - Redirect. Abaxis Medical Diagnostics, 2006. Web. 12 Sept.
2010. <http://www.abaxis.com/veterinary/vetscan_hm2_technology.html>.
Graham, Marshall Don. "The Coulter Principle: Foundation of an Industry."
The Association for Laboratory Automation :: Home. JALA Volume 8, Issue
6, Dec. 2003. Web. 12 Sept. 2010. <http://labautomation.org>.
Kelly, Kathleen. "Modules :: CLIA :: CME." Hematology in the Physician
Office Laboratory Section I. University of Iowa Carver College of Medicine,
2008. Web. 12 Sept. 2010.
<http://www.medicine.uiowa.edu/cme/clia/modules.asp?testID=4#02>.
Krantz, Alexander. "Residency & Fellowship Programs | Education."
Department of Pathology & Cell Biology | CUMC. Columbia University of
Physicians and Surgeons. Web. 12 Sept. 2010.
<http://pathology.columbia.edu/education/residency>.
Harmening., Denise, Clinical Hematology and Fundamentals of Hemostasis,
3rd edition, pp. 593-599.
Turgeon, Mary Louise, Clinical Hematology - Theories and Procedures, 3rd
edition, pp373, 376-382.
Rodak, Bernadette, Diagnostic Hematology, 1st edition, p.605-606.
Coulter STKS Operating Manual
McKenzie, Shirlyn, Clinical Laboratory Hematology, 2nd edition,pp 813-829.

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Electrical Impedance

Hemacytometer blood counts

In 1953, Wallace Coulter patented the Coulter

The Coulter Principle

Electrical Impedance (The Coulter

Sweep Flow: Steady stream of diluent that flows behind each

Most common chambers using impedance:

Cell the instrument

2-20 femtoliters (fL)

Red Blood Cell

RBC and Platelet

The black line represents normal cell distribution. The

Coincidence: Multiple cells passed through

In some analyzers the WBC count is directly

"Abaxis Veterinary Diagnostics - VetScan HM2 Technology." Abaxis Medical

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