COMPLETE BLOOD COUNT
LECTURE GUIDE
Prepared by: Rene Jesus Alfredo R. Dinglasan, RMT
ETHYLENEDIAMINETETRAACETIC ACID
EDTA (ethylenediaminetetraacetic acid)
 Found in: _____________________
 No. of inversions: ________
 Used for: ______________________
 M ost commonly used blood collection
tube in the Hematology section
 COMPLETE BLOOD COUNT (CBC)
 Also called: __________________
 Performed on automated
hematology profiling instruments
and includes:
RBC Parameters:
-RBC count
-Hemoglobin
-Hematocrit
-RBC indices
-Red Cell Distribution Width
(RDW)
-Reticulocyte count
WBC Parameters
-WBC count
-NEUT count: % and absolute
-LYM PH count: % and absolute
-M ONO count % and absolute
-EO and BASO counts: % and
absolute
Platelet Parameters
-Platelet count
-M ean Platelet Volume (M PV)
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-optimal anticoagulant concentration is: ________
-action: __________
-Important reminders in using EDTA:
●CBC can be performed on blood stored at room temperature for up
to 4 hours
●WBC counts, Hematocrit, and Platelet counts can be
determined up to 24 hours after blood is collected in
EDTA if it is refrigerated at 4 C.
[ *Delta Check – a process in which a current test result
is compared with the result of the same test from the previous
specimen from the same patient]
●ESR – should be set up within 2 hours of collection if the
EDTA blood was stored at room temperature or within
6 hours if the specimen was refrigerated
●Blood smears – can be made from EDTA tube as long as they
are made within 2 hours of blood collection
●EDTA prevents platelet aggregation (therefore the preferred
anticoagulant for platelet counts)
● Mean Platelet Volume (MPV)
- EDTA causes swelling of platelets (causes approximately 20%
increase in M PV during the first hour)
- Should be based on EDTA specimens that are between 1 to 4
hours old
●Insufficient EDTA
cause: ______________________
effect: ______________________
●Excessive EDTA
cause:________________________
effects: ________________________
HEMOGLOBIN and HEMATOCRIT

●HEMOGLOBINOMETRY (HEMOGLOBIN DETERMINATION)

◙Reference method: Cyanmethemoglobin (HiCN) Method
 Principle: Hemoglobin (Fe2+) + K3Fe (CN) 6  methemoglobin (Fe3+) + KCN  cyanmethemoglobin
 Uses Drabkin’s reagent - major components:
•Potassium ferricyanide (K3Fe(CN) 6) •Potassium cyanide (KCN)

 HiCN is measured at: ___________ (All types of Hb may be measured through this method, EXCEPT: __________________.)

 Possible sources of error:

SOURCES OF ERROR (FALSELY ELEVATED Hb) CORRECTION
High WBC count Centrifuge reagent-sample solution, then the supernatant is
measured
High platelet count

Lipemia Add 0.01 mL of the patient’s plasma to 5 mL of the cyanmet-

Cells containing HB S and HB C
Abnormal globulins (such as those found
in patients with plasma cell myeloma or
Waldenstrom macroglobulinemia) may precipitate in the
reagent.
hemoglobin reagent and using this solution as the reagent
blank
Make a 1:2 dilution with distilled water
(1 part diluted sample plus 1 part water) and multiplying the
results from the standard curve by 2.
Add 0.1 gram of potassium carbonate to the cyanmethemo-
globin reagent. Commercially available cyanmethemoglobin
reagent has been modified to contain KH 2PO4 salt, so this
problem is NOT likely to occur.

 Reminders:
o Cyanmethemoglobin reagent is sensitive to light (should be stored in a brown bottle or in a dark place)
o Another technique that has been used in some automated instruments involves the use of sodium lauryl sulfate (SLS)
to transform hemoglobin to SLS-methemoglobin. This method does NOT produce toxic wastes.
o HemoCue –an example of a commercially available handheld system to measure the hemoglobin concentration
- In here, hemoglobin is converted to azidemethemoglobin and is read photometrically at two wavelengths
(570 nm and 880 nm).

HEMATOCRIT DETERMINATION
-Reference ranges:
CONVENTIONAL UNITS S.I. UNITS
Adult Males 40 to 54% 0.40 to 0.54 L/L
Adult Females 35 to 49% 0.35 to 0.49 L/L
Newborn 53 to 65% 0.53 to 0.65 L/L

-“Hematocrit”- actually pertains to the instrument used to determine packed cell volume [PCV]
-After centrifugation of an anticoagulated whole blood specimen, the red blood cells along with other formed elements
(white blood cells and platelets) will settle at the bottom of the tube.
-The volume of the red blood cells that have settled is called the packed cell volume otherwise known as the hematocrit.
-Parallax: object being seen in a diff. position by changing the position of the head of the observer, or as seen by one
eye versus the other eye.
-Micro-Hematocrit Tube:
o Approx. 75 mm long
o Inner bore: 1.2 mm
o Can hold 0.05 mL of bld.

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 o Plug: 4 to 6 mm long (seal the capillary tubes at the end of the tube with the colored ring)
o 2 TYPES:
 ________________________

 ________________________

-Summarized procedure (using non-anticoagulated whole blood):

1.) Perform skin puncture.
2.) Wipe off the first drop of blood.
3.) Fill two heparinized capillary tubes two-thirds with blood.
(Note: Air bubbles denote poor skills but do not actually affect the test results).
(Tubes with a colored ring at one end are filled from opposite end.)
4.) In a vertical position, carefully seal the dry end of the heparinized capillary tubes with the sealing clay and the plug
should be 4 to 6 mm long.
5.) Place the two heparinized capillary tubes in the radial grooves of the microcentrifuge with their heads exactly opposite
each other. The sealed end should be away from the center of the centrifuge.
6.) Spin for 5 minutes at 10,000 RPM.
(Note: RPM must be checked periodically with a tachometer.)
7.) After centrifugation, read the hematocrit (the buffy coat layer should NOT be included).
(Note: Results should agree within + 0.02 L/L for the 2 patient samples run.)

-REMINDERS:
o Trapped plasma may cause the Hct to be falsely increased by as much as 0.02 L/L.
o When determined by fully automated methods, the Hct may be 0.01 to 0.03 L/L lower than the microhematocrit method
because it is electronically calculated and therefore is unaffected by trapped plasma. The difference in the hematocr it
results is usually insignificant, unless there is a more severe case of poikilocytosis and anisocytosis.
o Automated hematocrit – a calculated value from RBC and MCV
o Trapped plasma – small amount of plasma that remains in the erythrocyte portion of the spun Hct even when proper
centrifugation is used.
o More trapped plasma in the following:
 Sickle cell anemia
 Hypochromic anemia
 Spherocytosis
 Macrocytosis
 Thalassemia
o Certain abnormal RBC shapes (for example: spherocytes and sickle cells) inhibit complete packing.
o Immediately after a blood loss, Hct is not a reliable estimate of the degree of anemia because plasma volume is
replaced faster than RBC volume, therefore causing a temporarily lower Hct.
o POTENTIAL CAUSES OF ERRORS:
FALSELY INCREASED HCT FALSELY DECREEASED HCT

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RELATED TOPICS:

A.) Rule of Three

- used for checking validity of test results

-works only on normocytic, normochromic specimens

B.) RBC Indices

●Mean Cell Volume (MCV)

o Average volume of an individual RBC Cases:
o Formula: • ↓ MCV - _________________

• Normal MCV - _______________

o R.R.: 80 to 100 femtoliters • ↑ MCV - _________________

●Mean Cell Hemoglobn (MCH)

o Average weight or amount of hemoglobin in an individual RBC
o MCH follows the MCV (smaller RBCs necessarily hold less hemoglobin; larger RBCs can hold more hemoglobin)
o Formula:

o R.R.: 26 to 32 picograms

●Mean Cell Hemoglobin Concentration (MCHC)

o Mean concentration of hemoglobin in the average RBC Cases:
o Formula: • ↓ MCHC - _________________

• Normal MCHC - _______________

o R.R.: 31 to 37 g/dL or % • ↑ MCHC - _________________

REMINDERS:
•RBCs cannot accommodate more Hb than 37 g/dL; therefore, a result greater than 37 g/dL should be recomputed, making
sure that all values were accurately measured and no interfering substances are present.
•MCH and MCHC have lost some clinical value (however, all RBC indices are valuable quality control tools and help in the
recognition of instrument malfunctions.

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WBC DIFFERENTIAL COUNT

WBC Differential Count

■Types:
A.) 100-cell differential
 Routinely performed

B.) 200-cell differential

 Performed in instances when the WBC count is >40 x 109/L to increase accuracy of results
 NOTE:
o 200-cell differential may also be performed in cases when:
 ________________
 ________________
 ________________
 ________________

o Results are then divided by 2.

o Indicate in the report that 200 WBCs were counted.

CASE:
●Male patient, 29 years old
● WBC count: 10, 000/µL

●WBC differential count results (100-cell Differential):

 N = 25
 L = 68 Perform 200-cell
What should the medtech do
 M= 2 differential on the same
 E= 4 next?
PBS.
 B= 1

●WBC differential count results (200-cell Differential):

 N = 50
 L = 136 What should the medtech do Divide each of the
 M= 4 next? values by 2.
 E= 8
 B= 2

C.) 300 or 400-cell differential

 Performed in instances when the WBC count is >100 x 109/L to increase accuracy of results

D.) 50-cell differential

 Performed in instances when the WBC count is <1.0 X 109/L
 Performed using a buffy coat smear
 Count 50 WBCs, then multiply the results by 2 to get a percentage

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■ Reported as:
o RELATIVE COUNT
-gives the number of specific WBC type per 100 WBCs
-not as informative as the absolute count
-formula:
No. of specific WBC type X 100 = Relative count (%)
100 WBCs counted

-Reference Ranges:
• Neutrophil = ________
• Lymphocyte = ________
• Monocyte = ________
• Eosinophil = ________
• Basophil = ________

o ABSOLUTE COUNT
-gives the number of specific WBC type per cubic millimeter of blood (mm 3)
-more informative than the relative count
-formula:
Relative count (%) X WBC count = Absolute count (per mm 3 of blood)

-Reference Ranges:
• Neutrophil = ________
• Lymphocyte = ________
• Monocyte = ________
• Eosinophil = ________
• Basophil = _________

o Sample problem:
 Patient’s WBC count = 10,000/mm 3
 WBC Differential count:
o Neutrophil = 25%
o Lymphocyte = 68%
o Monocyte = 2%
o Eosinophil = 4%
o Basophil = 1%

 Interpretation: _____________________________

_____________________________

_____________________________

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EXAMPLES OF HEMATOLOGY ANALYZERS

Parameter Beckman Coulter Sysmex XN Series Abbott CELL-DYN Siemens ADVIA

Unicel DxH 800 Sapphire 2120i
WBC Impedance volume Fluorescent staining; Light scatter (primary Light scatter and absorp
and conductivity and forward light scatter count), impedance -tion
five-angle light scatter and side fluorescent (secondary count)
measurement light detection
RBC Impedance Impedance Impedance Low-angle
and high-angle laser
light scatter
HGB Modified cyanmet Sodium lauryl sulfate Modified cyanmethemo Modified cyanmethe-
hemoglobin (525 nm) -HGB (555 nm) globin (540 nm) moglobin (546 nm)

HCT (RBC X MCV)/10 Cumulative RBC pulse (RBC x MCV)/10 (RBC x MCV)/10
height detection
MCV Mean of RBC volume (HCT/RBC) x 10 Mean of RBC volume Mean of RBC volume
distribution histogram distribution histogram distribution histogram
MCH (HGB/RBC) x 10 (HGB/RBC) x 10 (HGB/RBC) x 10 (HGB/RBC) x 10
MCHC (HGB/HCT) x 100 (HGB/HCT) x 100 (HGB/HCT) x 100 (HGB/HCT) x 100
Platelet Count Impedance volume and Impedance; light scatter Dual angle light scatter Low-angle and high-
conductivity and ; fluorescent staining, analysis; impedance angle light scatter;
five-angle light scatter forward light scatter, count for verification; refractive index
measurement and side fluorescent optional CD61 mono-
light detection clonal antibody count
RDW RDW as CV (% ) of RDW-SD (fL) or Relative value, CV (% ) of RBC
RBC histogram or RDW-CV(% ) equivalent to CV histogram
RDW-SD (fL)
Reticulocyte Supravital staining; Fluorescent staining; Fluorescent staining; Supravital staining
Count impedance volume forward light scatter low-angle scatter, and (oxazine 750); low-
and conductivity and and side fluorescent fluorescent light detec- angle and high-angle
light scatter measure light detection tion light scatter and absor-
-ment bance
NRBC Impedance volume and Fluorescent staining; Red fluorescent dye Multi-angle light scatter
conductivity and five forward light scatter staining; forward light measurements in the
-angle light scatter and side fluorescent scatter and fluorescent two WBC differential
measurement light detection light detection channels
WBC Differential Impedance volume and Fluorescent staining; Multi-angle polarized Peroxidase staining,
conductivity and five forward and side light scatter separation light scatter and absorp-
-angle light scatter scatter, and side (MAPSS) and three tion;
measurement fluorescent light detec- -color fluorescence
tion detection for basos, differen-
tial lysis, low-angle and
high-angle laser light
scatter

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REFERENCE RANGES

TESTS REFERENCE RANGES**

RBC count Males = 4.6 to 6.0 X 1012/L
Females = 4.0 to 5.4 X 10 12/L
Red Cell Distribution Width (RDW) 11.5 to 14.5 %
Reticulocyte Count 0.5 to 1.5%
Hemoglobin Males = 14-18 g/dL
Females = 12-15 g/dL
Hematocrit Males: 40 to 54%
Females: 35 to 49%
Modified Westergren:
• Males: 0 to 10 mm/hour
Erythrocyte Sedimentation Rate
• Females: 0 to 15 mm/hour
Wintrobe:
• Males: 0 to 9 mm/hour
• Females: 0 to 20 mm/hour
Mean Corpuscular Volume (MCV) 80 to 100 fL
Mean Corpuscular Hemoglobin (MCH) 26 to 32 pg
Mean Corpuscular Hemoglobin 31 to 37 g/dL or %
Concentration (MCHC)
WBC count For both males & females:
4.5 to 11.5 x 109/L
WBC Differential Count (Relative Count)
Neutrophils 51 to 67 %
Lymphocytes 25 to 33%
Eosinophils 1 to 4%
Monocytes 2 to 6%
Basophils 0 to 1%
Platelet count 150 to 450 x 109/L
**Reference ranges are for adults using frequently-used units.
**The difference between 1mm 3 and 1 uL is INSIGNIFICANT .

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CASE STUDY
(NOTE: This case was based from “Quick Review Q and A’s” by: VALERIE DIETZ POLANSKY, M.
Ed., MT(ASCP))

A 40-year-old man was seen by his physician and he complained of shortness of breath, coughing, and fever. Interpret
the results of his hemogram.

RESULTS REFERENCE RANGES

WBC count 20.8 For both males & females:
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4.5 to 11.5 x 10 /L
12
RBC count 2.11 Males = 4.6 to 6.0 X 10 /L
12
Females = 4.0 to 5.4 X 10 /L
Hemoglobin 10.5 Males = 14-18 g/dL
Females = 12-15 g/dL
Hematocrit 24.5 Males: 40 to 54%
Females: 35 to 49%
RBC Indices
MCV 129.1 80 to 100 fL
MCH 52.8 26 to 32 pg
MCHC 41.2 31 to 37 g/dL
RDW 18.6 11.5 to 14.5 %
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PLT 232 150 to 450 x 10 /L

◆INTERPRETATION: ________________________________________________________________________

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●REFERENCES: Kindly refer to the Hematology Compiled Review Notes and Lecture Guide by: RJAR Dinglasan

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