Checklist 1: Performing white blood cells differential count

Skills

Not Performed

Not Applicable
Performed
Comments
Standard Criteria

1.1 Performing 1.1.1 wears of gloves

white blood
cells 1.1.2 Prepares all the requirements needed for white blood
differential cells differential count.
count 1.1.3 Prepares thin blood film

1.1.4 Stains with Leishman stain

1.1.5 Places a well stained slide on the stage of the

microscope, smear side up and focus using the low
power objective. (X10)
1.1.6 Checks to see if there are good counting areas
available free of ragged edges and cell clumps
1.1.7 Checks the WBC distribution over the smear

1.1.8 Checks that the slide is properly stained

1.1.9 Places a drop of immersion oil on the slide and

change the objective to X100 oil.
1.1.10 Reads and compute differential count of the white
blood cell
Checklist 2: Perform total white blood cell count using Turk’s solution
Skills

Not Performed

Not Applicable
Performed
Comments
Standard Criteria

2.1 Perform total 2.1.1 Wears PPE

white blood cell
count using 2.1.2 Mixes the blood sample carefully and add 0.38mL
Turk’s solution, of WBC diluting fluid in to a clean glass tube.
2.1.3 Adds 20μL of blood and mix it well. Leave it for
3-4 min.
2.1.4 Assembles the counting chamber and charge it
carefully with above suspension.
2.1.5 Focuses the ruling area of counting chamber under
10X look for even distribution of cells.
2.1.6 Counts the WBCs in the four larger outer square
using10X objectives and lower the condenser to
reduce the light intensity.
2.1.7 Useses formula to calculate total WBC count per
Number of cells counted
liter of blood= X dilution factor (20) X106
volume (4X1X1X0.1mm3)
2.1.8 Reports and interpret results
Checklist 3: Stain thin blood film to identify normal RBC morphology

Not Performed
Skills

Not Applicable
Performed
Comments
Standard Criteria

3.1 Stain thin 3.1.1 Wears PPE

blood film 3.1.2 Prepares a thin blood smear on a clean and dry microscopic
to identify glass slide and air dry it.
normal 3.1.3 Covers a well dried, thin blood smear with undiluted Leishman
RBC Stain solution by counting drops of Leishman stain.
morpholo 3.1.4 Lets to stand for 2 minutes, the methanol present in the stain
gy fixes the smear onto the glass slide.
3.1.5 Adds twice the amount of distilled water or Phosphate buffer
solution and mix the content by swirling or by blowing gently.
3.1.6 Incubates the slides for at least 10 min at 37 °C. This will stain
the blood cells.
3.1.7 Rinses the slides thoroughly with Phosphate buffer solution up
to 2 minutes or until it acquires a purple-pinkish tinge.
3.1.8 Air dries the slides in a tilted position so that the water easily
remove out of the slides.
3.1.9 Observes the slides under oil immersion objective lens of the
microscope.
3.1.10 Results interpretation
Checklist 4: Perform Reticulocyte count

Skills

Not Applicable
Not performed
performed
Comments
Standard Criteria

4.1 Perform 4.1.1 Wears of PPE

Reticulocyte
count using 4.1.2 Adds two drops of blood to three drops of NMB
New Methylene solution
Blue stain
4.1.3 Adds one more drop of blood if Hb is low.

4.1.4 Mixes and leave it for 15 minutes.

4.1.5 Incubates the mixture for 10 minutes at 37 °C.

4.1.6 Resuspends the cells before making the slides.

4.1.7 Makes three thin smears and let them dry.

4.1.8 Counts a total of 1000 RBCs under the 100 x oil

immersion lens.

4.1.9 Tries to make a thin smear with around 100 cells per
field.

4.1.10 Keeps the record of reticulocytes as well.

4.1.11 Reticulocytes appear greenish-blue and have

different concentrations of reticulum formation.

4.1.12 Counts the reticulocytes on all three slides and

compare the distribution.

4.1.13 Using the formula to calculate retics

Reticulocytes 100%= Total reticulocytes
1000 RBCs X 100
4.2 Perform 4.2.1 Wears of PPE
Reticulocyte
4.2.2 Mixes the blood with this stain and incubate for 15 to
count using
30 minutes.
Special stain
with brilliant 4.2.3 Makes the smear from this blood sample.
cresyl blue
4.2.4 Uses formula calculate the percentage of
reticulocytes cells.
 Skills

Not Applicable
Not performed
performed
Comments
Standard Criteria

4.2.5 Reports and interpret results

Checklist 5: Performing Bleeding time test

Skills

Not Performed

Not Applicable
Performed
Comments
Standard Criteria

5.1 Performing 5.1.1 Wears of PPE.

Bleeding time
test 5.1.2 Applies the blood pressure cuff to the arm just
above the elbow.

5.1.3 Inflates the device to 40 mm of mercury and

maintain at this level.

5.1.4 Cleans the anterior surface of the fore arm with

70% alcohol swabs.

5.1.5 Makes two clean punctures about 2mm long and

2mm deep being careful to avoid underlying vein.
5.1.6 Blots the blood with the filter paper every 15
seconds.
5.1.7 Starts the stop watch as the first drop of the blood
appears.
5.1.8 Stops the timer immediately when blood ceases
and record the time
5.1.9 Calculates the average of the 2 punctures and
record bleeding time.
5.1.10 Reports and interpret results
Checklist 6: Perform clotting time test

Not Performed
Not Applicable
Skills

Performed
Comments
Standard Criteria

6.1 Perform clotting 6.1.1 Wears PPE

time test by 6.1.2 Pricks the finger with the lancet
Capillary Method
6.1.3 Holds the capillary over the blood to fill the capillary
automatically
6.1.4 Breaks the capillary after regular intervals
6.1.5 Records the result when the clot start to form.
6.2 Perform clotting 6.2.1 Wears of PPE
time test by Test
Tube Method 6.2.2 Performs this test at 37 ° C.
6.2.3 Takes 4 ml of blood for the tube method and start the
time
6.2.4 Notes the time when there is the first appearance of the
clot formation
6.2.5 Uses multiple tubes
6.2.6 Records the results
6.3 Perform clotting 6.3.1 Wears of PPE
time test using Lee6.3.2 Uses two siliconized tube with 10 cm external bore
and White Method
6.3.3 Prewarms tubes at 37 °C in water bath
6.3.4 Takes blood sample from the antecubital vein.
6.3.5 Takes 2 to 2.5 mL of the blood and 1mL of the blood
is in each test tube
6.3.6 Starts two stopwatches as you see the blood in the
syringe
6.3.7 Keeps the blood in the water bath and check for
clotting by tilting each tube at 30 to 60 seconds
intervals.
6.3.8 Tilts the tube to greater than 90 degrees.
6.3.9 Stops the stopwatch as you see the clot in the tube.
6.3.10 Expresses the clotting time as the mean of the two
stopwatches.

6.3.11 Records and report results

Checklist 7: Performing Prothrombin time

Skills

Not Performed

Not Applicable
Performed
Comments
Standard Criteria

7.1 Performing 7.1.1 Wears of PPE

Prothrombin
time test 7.1.2 Collects the patient plasma and control plasma in
Sodium citrate tube

7.1.3 Adds 0.1ml of plasma into a glass tube and it should

be kept in a water bath at 37 Celsius

7.1.4 Adds 0.1ml of Thromboplastin into this tube

7.1.5 Waits for 1-3 minutes to allow the mixture to warm

7.1.6 Adds 0.1ml of CaCl2 and start the stopwatch

7.1.7 Stops the watch when the first fibrin appears

7.1.8 Carries out the test in duplicate on the patient’s

plasma and the control plasma

7.1.9 Reports the PT in seconds and interpret results

Checklist 8: Performing Activated partial Thromboplastin time

Not Performed
Not Applicable
Skills

Performed
Comments
Standard Criteria

8.1 Performing 8.1.1 Wears of PPE

Activated 8.1.2 Puts 0.1ml of plasma sample on the 2 test tube
partial labelled test 1 and test 2
Thromboplasti 8.1.3 Adds 0.1 ml of control plasma in 2 control tubes
n time test labelled control 1 and control 2
8.1.4 Adds 0.1 ml of APTT reagent to the 1 first tube (test 1
and control 1)
8.1.5 Incubates at 37 °C; stop watch is started as soon as the
reagent is added.
8.1.6 Incubates the tube labelled test 2 and control 2 one
minute after test 1 and control 1
8.1.7 Adds 0.1 ml of prewarmed CaCl2 & start the stop
watch.
8.1.8 Mixes the tube gently in the water bath for 20
seconds.
8.1.9 Takes the tube out & observe for clot.
8.1.10 Takes the average of the 2 readings.
8.1.11 Records and interpret results
Checklist 9: Perform Methaemoglobin reduction test (G6PD)

Not Applicable
Not Observed
Skills

Observed
Comment
s
Standard Criteria

9.1 Perform 9.1.1 Wears of PPE

Methaemoglo 9.1.2 Adds 0.1 ml of combined nitrate dextrose in the 0.1ml
bin reduction of blood sample followed by 0.1 ml of methylene blue.
test (G6PD) 9.1.3 Mixes the contents to brown colour
9.1.4 Incubates at 37 degrees for 3 hours
9.1.5 Transfers 0.1 ml of the incubated content in 10 ml of
distilled water
9.1.6 Incubates at R.T. for 10 minutes for colour development
9.1.7 Reports and interpret results
Checklist 10: Perform Eosinophil count

Not Performed

Not Applicable
Skills

Performed
Comments
Standard Criteria

10.1Perform 10.1.1 Wears of PPE

Eosinophil count 10.1.2 Dilutes blood sample in the WBC pipette as 1:
20 dilutions with staining fluids.
10.1.3 Mixes well for 30 seconds.
10.1.4 Leaves the chamber for at least 10 minutes to
count the cells.
10.1.5 Fills the chamber and count the coloured
Eosinophil.
10.1.6 Calculates and record results
Checklist 11: Prepare sensitized red blood cells according to SOP

Not Performed
Not Applicable
Skills

Performed
Comments
Standard Criteria

11.1Preparation of 11.1.1 Wears of PPE

sensitized red 11.1.2 Pools blood group O positive
blood cells 11.1.3 Washes the cells.
according to 11.1.4 Mixes the cells with incomplete Anti D (IgG)
SOP 11.1.5 Incubates the cells for 30 minutes at 37°C
11.1.6 Washes the cell.
Checklist 12: Perform Direct Coomb’s Test

Not Performed
Not Applicable
Skills

Performed
Comments
Standard Criteria

12.1 Perform 12.1.1 Wears of PPE

Direct 12.1.2 Prepares 5% cell saline suspension of the cells to be
Coomb’s Test tested.
12.1.3 Labels 3 tubes as T, PC and NC.
12.1.4 In the tube labelled as T (Test), take 2 drops of 5% saline
cell suspension to be tested.
12.1.5 Takes 1 drop of PC (Positive control), anti D sera and 1
drop of Rh +ve pooled cells.
12.1.6 Takes 1 drop NC (Negative control), of normal saline
and one drop of Rh +ve pooled cells.
12.1.7 Adds 2 drops of Anti human globulin to each of the
tubes.
12.1.8 Mixes well and centrifuge for 1 minute at 1500 rpm.
12.1.9 Resuspends the cells by gentle agitation and examine
macroscopically and microscopically for agglutination.

Checklist 13: Perform Indirect Coomb’s Test, Interpret, and comment on results

Not Performed
Not Applicable
Skills

Performed
Comments
Standard Criteria

13.1 Perform 13.1.1 Wears of gloves

Indirect 13.1.2 Labels 3 tubes as T, PC and NC.
Coomb’s Test 13.1.3 Takes 2 drops of test serum in the tube labelled T (Test)
13.1.4 Takes 1 drop of anti D serum in the test tube labelled
as PC (Positive control),
13.1.5 Takes 1 drop of normal saline in the test tube labelled
as NC (Negative control),
13.1.6 Adds one drop of 5% cell saline suspension of pooled O
Rh +ve cells in each tubes.
13.1.7 Incubates all the tubes at 37°C for 1 hour.
13.1.8 Washes the cells 3 times with normal saline.
13.1.9 Adds 2 drops of Anti Human Globulin to each tube.
13.1.10 Keeps for 5 minutes and centrifuge at 1500 rpm for 1
minute.
13.1.11 Resuspends the cells and examine macroscopically as
well as microscopically for agglutination.