CLINICAL HEMATOLOGY 1 (HEMA 311)

LECTURE NO. 18: INSTRUMENTATION, QUALITY CONTROL, AND OTHER ADDITIONAL NOTES
Prepared by: Roiland A. Baybayon, RMT, DTA, MLS (ASCPi), MSMLS(c)
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INSTRUMENTATION AND QUALITY CONTROL IN THE HEMATOLOGY LABORATORY
• Instrumentation and automation were first introduced by Joseph and Wallace Coulter
(Coulter Brothers) when they developed ___________________________________
COULTER COUNTER (1950’s),
the first type of automated equipment in the hematology laboratory

ADVANTAGES AND DISADVANTAGES OF AUTOMATION

ADVANTAGES DISADVANTAGES
✓ FASTER TURN-AROUND TIME ▪ COSTLY
✓ REQUIRES SMALLER SAMPLE VOLUME ▪ REQUIRES PREVENTIVE MAINTAINANCE
✓ INCREASED WORK-LOAD EFFICIENCY
✓ TESTS CAN BE ANALYZED THROUGH BATCH
TESTING

BASIC COMPONENTS OF MOST HEMATOLOGY ANALYZERS

✓ HYDRAULICS: aspirating units, dispensers, dilutors, mixing chambers, aperture baths, and
hemoglobinometer
✓ PNEUMATICS: vacuums and pressure for operating valves and moving sample through hydraulics system
✓ ELECTRICAL SYSTEMS: electronic analyzers and computing circuitry for processing data generated
NOTE: SCAN QR CODE TO
GENERAL PRINCIPLES OF AUTOMATED BLOOD CELL ANALYZERS: ACCESS VIDEO EXPLANATION OF
✓ ELECTRONIC IMPEDANCE ELECTRONIC IMPEDANCE ON
✓ OPTICAL SCATTER/ OPTICAL DETECTION YOUTUBE
✓ RADIOFREQUENCY
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A. ELECTRONIC IMPEDANCE/ELECTRICAL IMPEDANCE (MOST COMMON METHODOLOGY USED)
B. OPTICAL SCATTER/ OPTICAL DETECTION IMPORTANT NOTE:
• Also known as LOW VOLTAGE DIRECT CURRENT IMPORTANT NOTES:
• Also known as HYDRODYNAMIC FOCUSING • The NUMBER OF PULSES
RESISTANCE
• The NUMBER OF PULSES • LASER light is used (LASER: LIGHT AMPLIFIED by GENERATED is DIRECTLY
• Originally developed by the Coulter Electronics
GENERATED is proportional STIMULATED EMISSION of RADIATION) PROPORTIONAL to the
• Can be referred to as the COULTER PRINCIPLE
to the NUMBER OF • NOTE: FLOW CYTOMETRY: uses LASER LIGHT: most NUMBER OF CELLS
• BASIS: Cell counting and cell sizing are based on
the detection and measurement of changes in PARTICLES/CELLS counted common light source used in flow cytometers
electrical impedance/resistance produced by because of the properties in INTENSITY, STABILITY, ANGLES OF LIGHT SCATTER
• The AMPLITUDE/MAGNITUDE (Height) and MONOCHROMATISM
particles as they passed through a small aperture
• PRINCIPLE: Particles such as blood cells are non- of the electrical pulse • PRINCIPLE: A diluted blood specimen passes in a ✓ FORWARD LIGHT SCATTER (0°):
conductive but are suspended in an electrically produced is proportional to steady stream through which a beam of laser light relates to VOLUME OF CELL
conductive diluent (SALINE). As the dilute the CELL VOLUME is focused. As each cell passes through the sensing ✓ FORWARD LOW-ANGLE LIGHT
zone of the flow cell, it scatters the focused light. SCATTER (2°-3°): relates to CELL
suspension of cells is drawn through the aperture,
HISTOGRAM: Scattered light is detected by a photodetector SIZE or CELL VOLUME
the passage of each individual cell momentarily
✓ FORWARD HIGH-ANGLE LIGHT
increases the impedance/resistance of the and converted into an electrical pulse.
X-AXIS: REPRESENTS CELL SIZE SCATTER (5-°15°): relates to
electrical path between the two submerged • The application of light scatter means that a single
REFRACTIVE INDEX
electrodes Y-AXIS: REPRESENTS RELATIVE cell passes across a laser light beam, the light will
✓ ORTHOGONAL LIGHT SCATTER
NUMBER OF CELLS be reflected and scattered.
(90°): relates to INTERNAL
COMPLEXITY

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 CLINICAL HEMATOLOGY 1 (HEMA 311)
LECTURE NO. 18: INSTRUMENTATION, QUALITY CONTROL, AND OTHER ADDITIONAL NOTES
Prepared by: Roiland A. Baybayon, RMT, DTA, MLS (ASCPi), MSMLS(c)
-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

NOTE: SCAN QR CODE TO ACCESS VIDEO

EXPLANATION OF OPTICAL SCATTER/ FLOW
CYTOMETRY ON YOUTUBE
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TYPES OF INSTRUMENTAL ERRORS
cell size
*FORWARD ANGLE: relates to ______________________________ 1. APERTURE PLUGS: causes POSITIVE ERRORS (MOST COMMON PROBLEM IN CELL COUNTING)
2. BUBBLES IN THE SAMPLE: causes POSITIVE ERRORS
cell granularity/ lobularity
*SIDE ANGLE: relates to ____________________________________ 3. EXTRANEOUS ELECTRICAL IMPULSES: causes POSITIVE ERRORS
4. EXCESSIVE LYSING OF RBC’s: causes NEGATIVE ERRORS
------------------------------------------------------------------------------------------------------------ 5. IMPROPER SETTING OF APERTURE/THRESHOLD: causes EITHER POSITIVE OR NEGATIVE ERRORS
C. RADIOFREQUENCY (RF) aka Alternating Current Resistance
IMPORTANT NOTES:
NOTE:
• High-voltage electromagnetic current is used to Mnemonics:
✓ RADIOFREQUENCY is
detect cell size, based on the cellular density • POSITIVE ERRORS: FALSELY INCREASES THE CELL COUNT/ RESULT "BEA" (+ errors)
DIRECTLY PROPORTIONAL to
• NEGATIVE ERRORS: FALSELY DECREASES THE CELL COUNT/ RESULT
the NUCLEAR SIZE and
DENSITY of a cell
ADDITIONAL NOTES:
✓ RADIOFREQUENCY/
3-PART DIFFERENTIAL: includes: GRANULOCYTES, MONOCYTES, and LYMPHOCYTES
CONDUCTIVITY is related to 5-PART DIFFERENTIAL: includes: NEUTROPHILS, LYMPHOCYTES, MONOCYTES, EOSINOPHILS,
the NC RATIO, NUCLEAR and BASOPHILS
DENSITY, and CYTOPLASMIC
GRANULATION ------------------------------------------------------------------------------------------------------------
BLOOD CELL HISTOGRAMS
A. RBC HISTOGRAM
• Can measure cells as small as 24fL; cells that are counted in 24-36fL are rejected as
RBCs (20-24fL)
• NOTE:
✓ IF RBC’s ARE LARGER THAN NORMAL; THE CURVE WILL SHIFT TO THE RIGHT
✓ IF RBC’S ARE SMALLER THAN NORMAL; THE CURVE WILL SHIFT TO THE LEFT

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 CLINICAL HEMATOLOGY 1 (HEMA 311)
LECTURE NO. 18: INSTRUMENTATION, QUALITY CONTROL, AND OTHER ADDITIONAL NOTES
Prepared by: Roiland A. Baybayon, RMT, DTA, MLS (ASCPi), MSMLS(c)
-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
B. WBC HISTOGRAM
• FIRST PEAK (45fL-90fL): small mononuclear population of cells (LYMPHOCYTES)
• SECOND PEAL (90fL-160fL): minor population of large mononuclear cells (MONOCYTES)
• THIRD PEAK (160fL-450fL): normal mature types of granulocytes (NEUTROPHILS,
EOSINOPHILS, and BASOPHILS)

C. PLATELET HISTOGRAM
• Platelets are counted if the volume falls between 2fL-20fL
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ERRORS CAUSED BY NATURE OF THE SPECIMEN

NATURE OF THE SPECIMEN EFFECT

GIANT PLATELETS May be counted as RBC’s or WBC’s
FRAGMENTS OF LEUKOCYTE CYTOPLASM May be counted as RBC’s or platelets
INCREASED NUMBER OF SCHISTOCYTES Makes accurate RBC and platelet
count impossible
AGGLUTINATION OF RBC’S, WBC’S, OR PLATELETS Will cause false-negative results for
each count
AGGLUTINATED RBC’S OR PLATELETS May cause false-positive leukocyte
counts
PLATELET SATELLITISM Falsely low platelet counts
LYSE-RESISTANT CELLS (SICKLE CELL, TARGET CELLS, May result in high WBC counts
HYPOCHROMIC CELLS)

--------------------------------END OF LECTURE IN CLINICAL HEMATOLOGY 1-------------------------

STAT (Shorter Turn Around Time

- Greek word: "statin" THANK YOU SO MUCH FOR YOUR TIME AND EFFORT!
- Immediate results I HOPE YOU’VE LEARNED A LOT ABOUT THE BLOOD!
GOOD LUCK ON YOUR FUTURE ENDEAVORS!
BE SAFE ALWAYS FUTURE RMT’S!
-SIR ROI