HEMATOLOGY 1.

STAT: 1 hour sizing

- a branch of medicine concerning - not applicable for ESR conducts aperture current, and
the study of blood, the blood- determination
rinses instrument components
forming organs, and blood
1. Beckman Coulter Ac.T 5 Diff between analyses.
diseases.
Analyzer
used to lyse blood cells and to
- In the medical field, it includes
> a fully automated hematology determine
the treatment of blood disorders
and malignancies including types analyzer providing a complete hemoglobin concentration
of hemophilia, leukemia,
lymphoma and sickle cell anemia. WBC five-part differential, which is Reagent System

EACMed Hematology Section determined simultaneously by the Reagent

offers: AV (Absorbance Cytochemistry 3. WBC Lyse
1. Complete Blood Count (CBC) and Volume) Technology and 4. Diff Fix
2. Coagulation Studies:
Prothrombin Time, Activated WBC/BASO methodologies 5 Diff Rinse
Partial Thromboplastin Time Small Sample Volume Function
(APTT) and D-dimer
3. Erythrocyte Sedimentation 1. CBC: 30 uL Used to lyse red blood cells for the
Rate (ESR) leukocyte count
2. CBC+Diff : 53 uL
4. Reticulocyte Count
and to differentiate poly-nuclear
5. Clotting Time & Bleeding Throughput
basophils.
Time
> up to 60 samples/hour
6. Malarial Smear Used to lyse erythrocyte, fix
7. Peripheral Blood Smear (PBS) Sample Stability leukocytes, and

What specimen? > CBC: 48 hours @ room temp. differentially stain granules of
monocytes,
TEST > Diff:
neutrophils, and eosinophils
Complete Blood Count (CBC) 24 hours @ room temp.
Used as a rinsing agent
Erythrocyte Sedimentation Rate Reagent System
(ESR) Ac.T 5 Diff Control Plus
Reagent
Reticulocyte Count > A hematology quality control
1. Diluent, 20 L material
Prothrombin Time
2. Hgb Lyse, used to monitor the performance
Activated Partial Thromboplastin
400 mL of
Time
EACMed Ac.T 5 Diff analyzers in conjunction
D-dimer
Function with the instrument's reagent
Clotting Time & Bleeding Time
system
Malarial Smear used for counting and
differentiating blood cells >
Peripheral Blood Smear (PBS)
dilutes whole-blood samples Available in three levels (low,
Hematology Section Process Flow normal,
stabilizes cell membrane for
Turn around time accurate counting and and high)

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> • a quantitative, automated Isotonic solution that dilutes the
sample
Open stability: 15 days hematology analyzer and
Stabilizes cell membranes
Storage: 2-8°C WBC differential counter for
V Conducts aperture current
Machine Linearity invitro diagnostic use in
Carries and focuses the VCS
Parameter clinical laboratory.
sample stream
Unit • Modes of operation:
Rinses the system between
Linearity Range 1. Primary mode samples

Difference 2. Disrupts erythrocytes

(Whichever is Greater) Secondary r Frees hemoglobin

WBC Primary mode V Reduces cellular debris

103/uL > V Converts hemoglobin to a stable

cyanide-containing
0.5 to 80.0 Closed-Vial or Automatic Mode
pigment.
‡ 02 or+3% >
3. Erythrolyse Il erythrocyte
RBC Autoloader uses the rocker bed
and sensors to move specimen lytic reagent (also called
106/uL
tube
PAK LYSE)
0.20 to 7.50
holders called cassettes from the
4.
‡ 0.05 or ‡ 2 % loading bay to a cap-baying
station, StabiLyse leukocyte
Platelet
then to the unloading bay. preservative (also called
Hemoglobin
Aspirates 185 uL of sample from PAK PRESERVE)
g/dL each tube
5.
103/uL Secondary mode
COULTER CLENZ
10 to 1.000 >
cleaning agent
‡ 10 or ‡ 6 % Open-Vial or Manual Mode
Function
2.5 to 23.0 >
Dilutes the sample for the diff
‡ 03 or+ 2% Aspirates 125 uL of sample form
Lyses red blood cells
Hematocrit each tube
V Reduces cellular debris
% Reagent System
Preserves white blood cells in their
11.6 to 55.0 Reagent
near-native state
56.0 to 62.4 1. ISOTON Ill diluent
for differentiation using VS
+ 2 or + 3 % 2. technology

‡ 5 or + 5 % LYSE Ill lytic reagent Cleans and rinses the diluter parts

Beckman Coulter HmX AL Analyzer Function Prevents protein buildup

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Eliminates routine apperture Machine Linearity • The cells are suspended in a
bleaching conductive diluent and are pulled
Parameter
through
HmX 5C Cell Control
WBC
the apperture by low vacuum; this
> material used for quality control
RBC creates a momentary increase in
of CBC
Hemoglobin resistance to the electric flow.
and DIFF parameters
MCV • The resistance creates a pulse
Three levels of control with
that is sensed and counted by the
varying Platelet
instrument as a particle; the
mixtures of normal and abnormal MPV amount of resistance (the
values: Normal, Abnormal 1 and EACMed amplitude of

Abnormal 2. Unit each pulse) is directly related to

the size (volume) of the particle
Storage: 2-8 ° C 103/uL
that produced it
> 106/uL
Measurement
Open Stability: 13 days or 13 g/dL
events ( Parameter
fl
whichever comes first White Blood Cell
103/uL
Latron Control and Primer Hemoglobin
fL
> Daily quality control check to Hematocrit
verify performance of Linearity Range
Segmenters
the VCS technology TTM 0.0-99 9
Lymphocytes
components in both the 0.0-7.00
Monocyte
DIFF and Retics modes. 0.0-25.0
Eosinophils
1. LATRON Primer 50-150.0
Basophils
> Contains a surfactant to remove 0.0-999
bubbles and EACMed
5.0-20.0
clean the sample tubing and To 109-09
pathway from the tip Coulter Principle
le Blood Cell
of the aspirate probe to and • The Coulter Principle is an
electronic method for counting loglobin
through the flow cell
and sizing latocrit
2. LATRON control
particles; it is based on the fact Ac.T 5 Diff Analyzer
> Contains specific size latex that cells, which are poor
particles with a conductors of HmX AL Analyzer
predetermined volume, a weak current, will interrupt the derived from BC Histogram
conductivity and laser current flow.
Coulter Principle
light scatter
Photometric

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Calculated: (RBC x MCV) / 10 a. D-Dimer 1.2. Run three levels of cell
controls (low, normal and high).
Impedance with hydrotocus b. von Willebrand Factor - Activity
Results of
and Antigen
Derived from WBC scatterplot
which must be within the range
C.
using VCS technology specified in the product insert.
Free Protein S
3. ACL ELITE PRO 1.3. Enter the sample ID.
4.
a fully automated 1.4. Press CBC/Diff to select the
Special tests desired analysis mode (CBC or
coagulation system for CBC/Diff).
a.
specific clinical use in The mode selected appears on the
LAC Screen and Confirm screen.
hemostasis laboratory for
b. 1.5. Gently mix the sample by
clotting, chromogenic,
Silica Clotting Time Screen and invertion for about 6-8 times.
special assay and Make sure the
Confirm
immunological testing. sample is not clotted.
SAMPLE TRAY
ACL ELITE PRO 1.6. Remove the cap of the sample
~ 40 samples
used to perform the following tub.
~ 0.5 and 2.0 sample cups
test/s 1.7. Present the sample to the
~ full draw tubes probe and press the aspirate
1. Coagulometric tests
REAGENT POSITIONS switch. The
a. Prothrombin Time
Up to 22 total positions LEDs flash during sample aspirat
1
1. COMPLETE BLOOD COUNT 1.8. When the red LED remains
b. Activated Partial illuminated. remove the tube from
(AUTOMATED)
Thromboplastin Time the
1. Ac.T 5 Diff Analyzer
C. probe. When the green LED
1.1. Turn the instrument ON. The remains illuminated, the
Thrombin Time
instrument performs the Startup instrument is
2. Absorbance tests routine.
ready for the next analysis.
a. The Startup must be successful or
passed before sample analysis 1.9. The sample results appear on
Antithrombin the screen.
shall start.
b. 1.10. Retrieve the results using the
STARTUP ACCEPTABLE VALUE: sample ID in the interface. Fill in
Protein C
the
- WBC: 0.3 x 103/L
C.
necessary data.
- RBC: 0.03 x 105/L
Plasminogen
1.11. Validate the results
- Hemoglobin: 0.3 g/dL
d
1.12. Print results.
- Platelet: 7.0 x 10¾/L
Fibrinogen
2. HmX AL Analyzer
I. COMPLETE BLOOD COUNT
Immunological tests (AUTOMATED) 2.1. Machine start up
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2.1.1. Turn on the machine by 22.1.9. Press the "Esc" key. 2.2.2.5. Press F3 (Secondary)
pressing the switch at the back of
2.2.1.10. Load the cassette into 2.2.2.6. Mix the specimen gently
the
load elevator of the machine. but thoroughly.
analyzer.
2.2.1.11. Click "Sample Analysis. 2.2.2.7. Remove the cap of the
2.1.2. Turn on the monitor for the sample tube.
2.2.1.12. Click "Run Samples"
Data Management System (DMS)
2.2.2.8. Immerse the aspirator tip
by 2.2.1.13. Press "Enter"
completely in the tube.
pressing the switch on the side of 2.2.1.14. Press F3 (Run).
2.2.2.9. Press and release the
the instrument.
2.2.1.15. Press F2 ( Start sample bar.
2.1.3. Check status of reagents.
2.2.2. Secondary Mode 2.2.2.10. Listen for the beep
2.1.4. Check the waste container. before removing the tube.
2.2.2.1. From the Main Menu,
2.1.5. On the Main Menu of the select "Sample Analysis. 2.3. Retrieve the result using the
DMS, select "Diluter Functions" sample ID in the interface. Fill in
then 2.2.2.2. Click "Run Samples" the

"Start Up" to begin machine start 2.2.2.3. Type the sample ID and necessary data.
up. It must be successful or patient name then press "Enter"
2.4. Validate the results.
passed before starting the analy key.
2.5. Print the results at the back of
2.2. Running of samples 2.2.2.4. Press F3 (Run) the transaction slip.

2.2.1. Primary Mode 2.2.2.5. Press F3 (Secondary) 2.2.2. Secondary Mode

2.2.1.1. Put the specimen tube 2.2.2.6. Mix the specimen gently 2.2.2.1. From the Main Menu,
into the cassette making sure any but thoroughly. select "Sample Analysis.

bar code labels are visible through 2.2.2.7. Remove the cap of the 2.2.2.2. Click "Run Samples"
the top of the sample tube.
2.2.2.3. Type the sample ID and
cassette. 2.2.2.8. Immerse the aspirator tip patient name then press "Enter"
completely in the tube.
2.2.1.2. From the Main Menu, key.
select "Sample Analysis." Click 2.2.2.9. Press and release the
sample bar. 2.2.2.4. Press F3 (Run)
"Worklist."
2.2.2.10. Listen for the beep 2.2.2.5. Press F3 (Secondary)
2.2.1.3. Press "F12" (Seq) before removing the tube.
2.2.2.6. Mix the specimen gently
2.2.1.4. Press the "Enter" key 2.2.2. Secondary Mode but thoroughly.
twice
2.2.2.1. From the Main Menu, 2.2.2.7. Remove the cap of the
2.2.1.5. Input the cassette number select "Sample Analysis. sample tube.
in use.
2.2.2.2. Click "Run Samples" 2.2.2.8. Immerse the aspirator tip
2.2.1.6. Press "F12" (Seg) completely in the tube.
2.2.2.3. Type the sample ID and
2.2.1.7. Press the "Ent patient name then press "Enter" 2.2.2.9. Press and release the
sample bar.
2.2.1.8. Input appropriate patient key-
identifiers (sample ID and 2.2.2.10. Listen for the beep
2.2.2.4. Press F3 (Run) before removing the tube.
patient name).
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2.3. Retrieve the result using the 1.7. When the red LED remains 2.3.5.2. Press and release the
sample ID in the interface. Fill in illuminated, remove the tube from sample bar.
the the
2.3.5.3. Listen for the beep or
necessary data. probe. When the green LED verify the status line message
remains illuminated, the
2.4. Validate the results. changes from "Aspirating" to
instrument is
"Diluting" before removing
2.5. Print the results at the back of
ready for the next analysis.
the transaction slip. the bottle.
1.8. Repeat steps 4 through 8 until
• RUNNING OF CELL CONTROL 2.3.6. After the analysis, review
you have run all three levels of cell
the primer results at the DMS.
1. Ac.T 5 Diff Analyzer
control.
2.3.6.1. If count is unacceptable,
1.1. Turn the instruments ON. The
1.9. Review the control results to rerun the Latron primer.
instrument performs the Startup
ensure they are within the
routine 2.3.6.2. If count is acceptable, run
acceptable
Latron Control.
The Startup must be successful or
ranges.
passed before sample analysis 2.4. Running of Latron Control
shall 1.10. Print quality control result.
2.4.1. Press the "Esc" ley to close
start. 1.11. Log the results in the the primer Run Window.
designated log she
1.2. Enter the cell control level 2.4.2. Press "F3" (Run).
(LOW. NORMAL OR HIGH) as the 2. Hm AL Analyzer
2.4.3. Press "F3" [Control(Seconda
sample
2.1. Ensure that start up is
2.4.4. Gently invert the Latron
ID. complete.
Control bottle 5 to 8 times,
1.3. Press CBC/Diff to select the 2.2. Bring Latron Primer and
2.4.5. Run the Latron Control in
desired analysis mode (CBC or Control at room temperature.
the Secondary mode.
CBC/Diff).
2.3. Running of Latron Primer
2.4.5.1. Immerse the aspirator tip
The mode selected appears on the
2.3.1. From the Main menu of the completely in the Primer. (Do
screen.
DMS software. select "Controls."
not place the tip against the side
1.4. Gently mix the sample by
2.3.2. Select "Control Run" of the bottle.)
inversion for about 6-8 times.
Inspect the 2.3.3. Press the "F2" key (File). 2.4.5.2. Press and release the
sample bar.
vial's contents to ensure that all 2.3.4. Select the current Latron
cells are uniformly distributed. File (active lot number) 2.4.5.3 .Listen for the beep or
verify the status line message
1.5. Remove the cap of the sample 235. Press "F3" (Run).
tube. changes from "Aspirating" to
2.3.4. Press F4 (Primer).
"Diluting" before removing
1.6. Present the sample to the
probe and press the aspirate 2.3.5. Run the Latron Primer in the
the bottle.
switch. The Secondary mode.
2.4.5.4. After the analysis, review
LEDs flash during sample 2.3.5.1. Immerse the aspirator tip
the the results in the Control
aspiration. completely in the Primer. (Do
Run Screen.
not place the tip against the side
of the botti 2.5. Preparation of 5C cell control
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2.5.1. Warm control vials at - Reconstitute with 8.5 mL PT 1.1. PT Fibrinogen HS Plus
ambient temperature for 10-15 Buffer (5 days upon
- Reconstitute with 8.5 mL PT
minutes reconstitution).
Buffer (5 days upon
EAGMed before mixing. 1.2. APTT Reagent Synthasil reconstitution).

2.5.2. Mix gently by hand using the - Liquid ready reagent and has 30 1.2. APTT Reagent Synthasil
&x8x8 method twice days stability upon opening.
- Liquid ready reagent and has 30
2.5.3. After the second mixing, 1.3. Normal Control Assayed days stability upon opening.
check the sides and bottom of the (Normal Control)
1.3. Normal Control Assayed
tube
- Reconstitute with 1 mL distilled (Normal Control)
to verify the control is mixed water (24 hour stability upon
- Reconstitute with 1 mL distilled
completely.
reconstitution). water (24 hour stability upon
2.6. Running of 5C Cell Control
1.4. Calibration Plasma (Normal reconstitution).
2.6.1. From the Main menu of the pool)
1.4. Calibration Plasma (Normal
DMS software. select "Controls.
- Reconstitute with 1 mL distilled pool)
2.6.2. Select "Control Run" water (24 hour stability upon
- Reconstitute with 1 mL distilled
2.6.3. Press the "F2" key (File). Reagent preparation water (24 hour stability upon

2.6.4. Select the current 5C Cell 1.1. PT Fibrinogen HS Plus reconstitutio

Control File (active lot number)
- Reconstitute with 8.5 mL PT 2. Machine start up
2.6.5. Run the 5C Cell Control in Buffer (5 days upon
2.1. Turn on the analyzer.
Primary Mode reconstitution).
2.2. Input username and password
2.6.6. Place the control tubes into 1.2. APTT Reagent Synthasil
in the monitor for Data
the cassette making sure any bar
- Liquid ready reagent and has 30 Management
code labels are visible through the days stability upon opening.
System (DMS).
top of the cassette.
1.3. Normal Control Assayed
3. Check status of reagents and
2.6.7. Load the cassette in the (Normal Control)
consumables.
autoloa
- Reconstitute with 1 mL distilled
4. Running of samples
2.6.8. Press F3 (Run). water (24 hour stability upon
4.1. On the home screen of the
2.6.9. Press F2 ( Start Primary). reconstitution).
DMS, click "Analysis"
2.7. Validate the results. 1.4. Calibration Plasma (Normal
4.2. Click "Multi-test session" or
pool)
2.8. Log the results in the "Single test session"
designated logsheet. - Reconstitute with 1 mL distilled
4.3. Click the box beside "Loadlist
water (24 hour stability upon
2.9. Remove the cassette from the No.
unload elevator of the mac reconstitu
4.4. Input load list number.
Il1. RUNNING OF SAMPLE (ACL FACMed
4.5. Place the sample in the
ELITE PRO)
Il1. RUNNING OF SAMPLE (ACL Sample tray.
1. Reagent preparation ELITE PRO)
4.6. Select the sample position.
1.1. PT Fibrinogen HS Plus 1. Reagent preparation
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4.7. Type the sample ID and the 3.3. Click "Loadlist No. Centrifuge at 3000G (for the
patient name. period of time recommended by
3.4. Input loadlist number.
the
4.8. Select the test's.
3.5. Place the control in the
manufacturer of the centrifuge-
4.9. Click the"" icon. Sample trav.
usually 10 minu
4.10. Click the "Run" icon to start. 3.6. Select the sample position.
V. ERYTHROCYTE VOLUME
5. After the analysis, validate the 3.7. Click "Add QC". FRACTION
results.
3.8. Select the test/s. (HEMATOCRIT)
6.
3.9. Click the "" icon. 4. Hold the tube against the scale
Transfer the result from the DMS so that the bottom of the column
3.10. Click the "Run" icon to start. of
to the interface by clicking the
"Host 4. Validate the results. erythrocytes (not the bottom of
Communication" icon in the lower 5. Click "QC" on the main screen the tube) is aligned with the
right side of the Database view horizontal
screen. 6. Log the results in the designated
quality control log sheet. zero line.
6.1. Click the box beside "Sample
7. Remove the sample from the 5. Move the tube across the scale
ID".
sample tray until the line marked 1.0 passes
6.2. Click "Start Communication"
V. ERYTHROCYTE VOLUME through the top of the plasma
7. Retrieve the result in the FRACTION column. Check to make sure that
interface. the
(HEMATOCRIT)
8. Fill in the necessary data. bottom of the column of
quality control result. erythrocyte is still on the zero line;
9. Print the results at the back of also check
the transaction slip. the results in the designated log
she (by means of also check (by means
10. Remove the sample from the of the heavy vertical lines) that
samp eMed.

1. Allow the blood to flow into the the tube is vertical.

IV. RUNNING OF CONTROLS (ACL
ELITE PRO) tube by capillary action. Fill about 6. The erythrocyte volume fraction
three quarters of a capillary tube reading is made exactly at the top
1. Reconstitute Normal Control of
Assayed (Hemosil) with one mL with blood from a finger puncture.
distilled water 2. Plug the other end of the tube, the column of erythro

(24 hour stability upon i.e., the end that has not come VI. LEUKOCYTE NUMBER
reconstitution). into CONCENTRATION

2. Check status of reagents. contact with the blood, with soft 1. Place a dampened gauze in the
wax or plastic modeling clay. bottom of petri dish with two
3. Running of control. Check halves of
3.1. On the main screen, click that it is completely plugged to a wooden stick placed on the gauze
"Analysis" depth of about 2mm. to serve as stand for the
3.2. Click "Multi-test session" or 3. hemocytometer.
"Single test session",

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2. Place a whole blood in a rotor remove and clean the coverslip; 1. Collect a drop of blood of about
for at least 1 to 2 minutes or hand clean the counting chamber; and 4mm in diameter on one end of
invert refill the

gently for at least 8 times. with another drop slides.

3. To make a 1:20 dilution, attach 11. Leave the counting chamber 2. Hold the slides with one hand.
a suction device to the pipette and on the bench for about 3 minutes Using the other hand, place the
draw a to allow edge of

sample of whole blood to exactly the cells to settle. Be careful it the spreader just in front of the
0.5 mark on pipette stem. Do not does not dry up drop of blood.
allow
12. Place the chamber on the 3. Draw the spreader back until it
air bubbles to enter the pipette. stage of the microscope. Use the x touches the drop of blood.
10
4. Tilt pipette so that the stem is 4. Let the blood run along the
slightly above horizontal and wipe objective. edge of the spreader.
stem
13. Count the leukocyte in area of 5. Push the spreader to the end of
carefully with slightly dampened 4mm3 of the chamber. Include in the slides with a smooth
gauze, moving from pipette bulb the count movement (all
toward
the leukocytes seen on the lines of the blood should be used).
tip two sides of each square counted,
6. Air dry smear. Adequate drying
5Gently rotating pipette between using the letter "L" or "." as your is essential to preserve the quality
forefinger and thumb, draw guide. This represents one of the of the
diluent four
smear.
steadily into pipettes exactly to squares counted. Repeat the
7. Write with a lead pencil on the
the 11 mark. procedure in the three other.
thick part of the smear not used
6. Immediately cover tip of pipette 14. Calculate the number of for
with a finger and remove the leukocytes in 1 liter of blood by
examination patient's name and
suction multiplying the
date of examination.
device. number of leukocytes counted in
8. Fix the thin smear with
the four squares by 0.05. Report
7. Shake pipette to mix by hand. methanol for 2-3 minutes.
the
8. 9. Stain according to the
result as "number" by 109
manufacturer's instructions.
Place a coverslip on the
Example:
hemocytometer. 10. Wash the stain off in a stream
Number of leukocytes counted in of buffered water. Do not tip the
9. Discard the first four drops.
4 squares = 188 stain off as
10. Dispense the mixture into the
Number of leukocytes per liter = thin will leave a deposit of stain on
counting chamber. If the liquid
(188 x 0.05) x 109 the smear.
overflows
Result Reported: 9.4 x 109 11. Leave clean water on the slide
into the channel between the two
for 2-3 minutes to differentiate
chambers, you must start again: VI. PREPARATION AND STAINING
the smear.
OF

BLOOD SMEAR
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12. Tip off the water and place the and never substitute other types 1. Using at least 30 specimens
slide in a draining rack to dry. of lubricants for immersion oil, from healthy subjects, perform
since machine
13.Examine the color of the
smear. It should appear mauve- substitutes may damage optical platelet count in a hematology
neither too blue lenses. analyzer. Record results.

nor too pink 4 2. Prepare smears from each

sample tested.
14. Check if the smear is Classify 100 leukocytes using
satisfactory. Characteristics of battlement track method. 3. For each of the 30 smears,
satisfactory count and record the number of
5.
platelets in
smear include the following;
Note down the total number for
10 consecutive fields. Use only the
14.1. There should be no lines each type.
area where 50% of the red cells
extending across or down
IX. PLATELET ESTIMATION are
through the smear.
A platelet estimate may be done overlapping in doublets or triplets.
14.2. The smear must be smooth on a smear to verify a
4. Divide the total number of
at the end, not ragged and questionable
platelets found by 10 to obtain the
lined. analvzer result. average

14.3. The smear must not be too 1. Count the number of number per single oil-immersion
long and too thick. identifiable platelets in 10 oil field
immersion fields by
14.4. The smear must not contain Platelet Estimation Factor
holes because a greasy slide using only the area where 50% of
5. For each specimen, divide the
the red cells are overlapping in
has been used corresponding machine count by
doublets or triplets. the
VIII.LEUKOCYTE DIFFERENTIAL
COUNT 2. Divide the total number of average number of platelets per
platelets counted by 10 to obtain oil-immersion field to obtain
1. Check slide identification. individual
the average
2. Perform low power scan to platelet estimation factors (PEF)
number per field.
review blood smear adequacy.
3. Multiply the average number by 6. Add all individual PEF from 30
2.1. Check feather edge for fibrin specimens and divide by 30 to
the platelet estimation factor.
clots. obtain the
4
2.2. Examine smear edges for average PEF. Round-off the
possible aggregation of The number obtained is reported number to the nearest digit and
leukocytes. as Platelet Estimate: use this
2.3. Verify stain quality x 109 number as the PEF.
2.4. Note also erythrocyte Platelet Estimation Factor Formula:
distribution pattern, size and
> derived by correlating a machine MC/SC = iPEF
shapes.
count and a smear count, as
3. Perform oil-immersion follows: PEF = Total iPEF ÷ 30
examination of the blood smear.
Legend:
Use oil sparingly
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MC = Machine Count oxalated vacuum tube. Mix well 6. At interval of 30 seconds, blot
but gently. the blood which has oozed out
SC = Smear Count
with
2.
¡PEF = Individual PEF
absorbent or filter paper.
Uncap the tube and insert gently
X. RETICULOCYTE COUNT
the commercially available 7. Record the time until the
1. Place three drops each of Westergren tube bleeding stop
reagent and blood sample in a test
unto the tube with venous tube. XIII.CLOTTING TIME
tube.
3. 1. Obtain four drops of blood by
2. Mix blood and reagent by
prick method and place them
swirling the tube. Check that the Westergren tube is
separately on
filled with blood up to the zero
3. Allow the blood and reagent
mark. a clean slide. Time simultaneously.
mixture to stand for 15-20
minutes. 4. 2. At intervals of one minute, draw
the tip of the needle to each of the
4. Allow the tube to stand in a
drops
vertical position in the Westergern
With the use of a pasteur pipette
rack exactly one successively. The appearance of
(or any of equivalent), place 1
fibin at the point of the needle is
drop of hour at room temperature
the
without vibration or exposure to
the mixture on one side of clean
direct sunlight. coaqulation time
slide.
5. 3. Record the results.
5. Prepare a thin blood dye film in
usual manner and allow to dry. After exactly 60 minutes or one CBC Reference Range
hour, the distance from the O
6. Using oil immersion lens, count Parameter
mark to the top of
the number of reticulocytes per
Unit
1000 the column of red cells is recorded
in milliliters as the ESR value. If the Screenshot (74).png
erythrocytes counted and record.
demarcation between plasma and Reference Range
7. Calculate the percent
the red cell column is hazy, the
erythrocytes using the following 2
level is taken
formula:
where the full density is firts appar NATIONAL EXTERNAL QUALITY
Number of reticulocyte counted y ASSESSMENT
1000 = Reticulocyte ct. in % XI1. BLEEDING TIME
SCHEME (NEQAS) ENROLLMENT
1000 RBC 1. Clean the ring finger of the
patient with cotton and alcohol. > aims to help laboratories attain
XI. ERYTHROCYTE accurate and reliable analytical
SEDIMENTATION RATE 2. Let it dry.
results and achieve inter-
(ESR) 3. Make a deep puncture on the laboratory comparability as well.
tip of the ring finger using sterile
1. > one of the requirements for the
lancet.
renewal of the License to
Collect venous blood into an EDTA
4. Discard the first drop
(Ethylene Diamine Tetraacetic Operate (LTO) of Clinical
Acid) tube or 5. Start timer. Laboratories of all category.

JMSE
> Reference Laboratory for
Hematology: National Kidney and

Transplant Institut

NEQAS Sample Processing and

Testing

1. Allow previously refrigerated

vials to stand at room
temperature (20-

24°C) for at least 15 minutes

before mixing.

2. Verify that the vial cap is

securely in place. Mix the vial by
gently end-to-

end inversion until the cell is

completely re-suspended.

3. Roll the vial gently between the

palms of the hands for 20 seconds
in the

upright position, invert vial and

roll 20 seconds more.

4. Process the samples using the

routine blood count analyis
method of

your Laboratory.

NEQAS Data Reporting

1. Report results according to the

units and decimal places specified
in the

Survey Result Form.

2. Fill up the Survey Result Form/s

completely and send back the
form to

the NRL either via email or

personal delivery

JMSE