Hematology-1
wbc
counting
sissay, aysha
WHITE BLOOD CELL COUNT
Denotes the number of white blood cells in 1 liter (L) of
whole blood.
Normal (healthy)-4,000 to 11,000 X 106/L (or 4.0 to 11.0
x 109/L)
Count varies with age
Newborn- 10.0 to 30.0 X 109/L at birth.
1 yr- 6.0 to 17.0 X 109/L
21 yrs- drops to normal levels
WHITE BLOOD CELL
Utilized to indicate infection
Employed for disease progression and therapies
Elevated WBC- bacterial infection, appendicitis, leukemia,
pregnancy, hemolytic disease of the newborn, uremia, and
ulcers.
Drop below NV- viral diseases (such as measles),
brucellosis, typhoid fever, infectious hepatitis, rheumatoid
arthritis, cirrhosis of the liver, and lupus erythematosus.
Radiation & drug therapys
WHITE BLOOD CELL
Leukocytosis-white blood cell count above 11.0 X 109/L
Leukopenia- a white blood cell count below normal
White count in children usually shows a greater variation in
disease.
Children with infections have higher WBC than adult’s
WBC variations:
Slightly higher in the afternoon than in the morning
Increase due to strenuous exercise, emotional stress,
anxiety
Laboratories- electronic counting of cells
GUESS THE
PICTURE
MANUAL WBC
COUNT
REAGENTS & EQUIPMENT
1. Pipets, one of the following:
a. WBC Unopette (1:20 dilution) is recommended because
of ease of use and technologist safety.
b. 20 mL pipet (also 10 X 75 mm test tubes with caps,
and plain microhematocrit tube if this pipet is used).
c. Thoma white count pipet.
REAGENTS & EQUIPMENT
2. Microscope.
3. Clean gauze or Kimwipes.
4. Improved Neubauer hemocytometer (counting chamber) with
coverglass.
The hemocytometer with Neubauer ruling hemocytometer
consists of two identically ruled platforms with raised ridges, a
cover glass, and a 0.1 mm space between the top and cover glass.
Two platforms have ruled areas of 9 mm large square with equal
size (3mm wide and 3mm long), with each square being 1 mm wide
and 1 mm long.
REAGENTS
&
EQUIPMENT
SPECIMEN
Whole blood, using EDTA as the anticoagulant.
Capillary blood may also be used.
PRINCIPLE
Whole blood is mixed with a weak acid solution
to dilute the blood and hemolyze the red blood
cells.
PRINCIPLE
Glacial acetic acid lyses the red cells.
Allowing a proper count of white cells.
Gentian violet slightly stains the nuclei of
the leucocytes.
PREPARATION OF DILUTED BLOOD
1. Aspirate blood up the 0.5 mark of the WBC pipette.
2. Wipe off the excess blood at the tip of the pipette with a
clean piece of tissue paper. Make sure that the tissue paper
does not touch the opening of the bore.
3. Place the diluting fluid in a clean transparent container, then
immerse the pipette. Use syringe aspiration to draw the
diluting fluid up to the 11 mark with constant rotation of the
pipette (3-5 minutes). This will ensure proper mixing of the
blood and the diluting fluid. This makes a 1:20 or 1/20
dilution.
FILLING THE COUNTING CHAMBER
1. Place the thick coverslip on top of the improved Neubauer
counting chamber. Both the coverslip and counting chamber must
be free from dirt, thumbmarks, tissue strands, and the like.
2. Reshake the pipette. Discard the first 2-3 drops of the
diluted blood from the pipette and carefully and exactly fill or
charge the counting chamber by capillary action. This can be
achieved by placing the tip of the pipette on the space between
the coverslip and the central platform of the counting chamber.
The angle of the pipette while charging is 30-35 degree
NOTE: Overcharging can be readily observed by the presence of
fluid on the moats of the counting chamber. Undercharging, on the
other hand, can be observed if the entire ruled area of the counting
chamber is not adequately covered. Air bubbles in the counting
chamber indicates the presence of moisture or dirt. If this occurs,
clean the coverslip and the counting chamber and repeat the process.
3. Let the chamber stand for 5-10 minutes to allow the WBCs to
settle.
4. Locate the ruled area for WBC counting by focusing the microscope
using the LPO.
PROCEDURE
Calculation of the WBC.
a. For each of the two white counts performed, calculate the number of
WBCs/uL, as shown below:
Number of WBC X Correction for Vol. X Correction for dilution
Number of white blood cells counted. Add the total number of WBC counted in
the four large squares of the counting chamber. For example:
Square 1 ------ 25 white cells ------ 25
Square 2 ------ 34 white cells ------ 34
Square 3 ------ 32 white cells ------ 32
Square 4 ------ 31 white cells ------ 31
Number of cells counted = 122
CORRECTION FOR DILUTION
Since the blood was initially diluted 1:20, the
correction factor for dilution is 20 (21 if using a
1:21 dilution).
PROCEDURE
b. Therefore:
WBC/uL = 122 X 2.5 X 20 = 6,100 WBC/uL
WBC/L = 6,100 X 10(6) = 6.1 X 10(7)/ L
d. Calculate the WBC for the second white count and
average the two numbers for the final result.
important
things to
consider
Diluting fluid must be free from contamination to prevent inaccuracies
and difficulties in counting white blood cells due to small blood amounts
often collecting.
The counting chamber and cover glass should be free from dirt and lint
to prevent inaccuracies and difficulties in counting white blood cells,
and should be cleaned off immediately after the count.
Pipets should be clean and free of dirt and dried blood, and undiluted
blood should not be left in a Thoma pipet as it can harden and plug the
pipet.
The hemocytometer must be filled promptly to avoid cell count
inaccuracies due to fluid evaporation in the chamber.
Thank You
mwah sana may na learn kau :))
