NAME : August 29, 2019
IMMUNOHEMA LAB 1 8:00 – 11:00 TH
Experiment Number 3
SAMPLE PREPARATION FOR BLOOD BANKING PROCEDURE
1. WHOLE BLOOD PREPARATION
A. Draw a sufficient amount of blood B. To achieve an optimum ratio of blood to
corresponding to the volume of the additive/ anticoagulant, the volume of
additive/ anticoagulant blood should
Fill the tube to the line indicated
on the evacuated tube label or
Be according to the appropriate
ratio of the sample to the
additive/ anticoagulant
c. Mix evacuated tube immediately after collection, C. Label the specimen container with patient
before clotting can occur, gently by inversion information including the date and time
according to the type of additive/ anticoagulant of collection
2. PLASMA PREPARATION
A. Draw sufficient amount of blood into an B. Gently mix immediately after collection
evacuated tube with the required by appropriate number of inversion
anticoagulant to yield the plasma volume according to anticoagulant used. Avoid
required for the test hemolysis of the specimen during
collection and mixing
C. centrifuge anticoagulated blood for 5-10 D. After centrifugation, 3 layers where
min. at 2500 rpm formed (from top to bottom) plasma,
buffy coat, red blood cells .
The plasma should appear clear and no pink to
red tinge is manifested.
E. Carefully aspirate or collect the
supernatant (plasma) with pasteur pipette
and transfer to test tube.
F. Label the sample container.
3. SERUM PREPARATION
A. Draw a sufficient amount of blood to B. Allow the tube to stand in upright
yield a serum volume required by the position at room temperature for 20-
test 30 mins.
C. After clotting incline the tube gently D. Centrifuge for 5-20 mins at 2500 rpm
and when it does not move,
coagulation time is reached. Then rim
the sides using applicator stick to depth
of 1 cm side to side.
E. Check the supernatant(serum) and
carefully aspirate it with a Pasteur
pipette and transfer to test tube .
Serum should appear clear and no
pink to red tinge is manifested.
F. Label the sample container.
4. RED CELL SUSPENSION PREPARATION
a. Transfer carefully ≤1.0 mL of the packed b. Washing phase
red blood cells from plasma preparation (1) Add normal saline solution (NSS)
into a graduated centrifuge tube up to the 10 mL mark of the
centrifuge tube
(2) Cover the centrifuge tube and
centrifuge the tube for 5-10 min at
2500 rpm then discard the
supernatant fluid using a pasteur
pipette, leaving the red blood cells
c. After final washing, note the volume of d. Computation
the washed packed red blood cells
before the supernatant is completely
removed by aspiration
e. Add the appropriate volume of NSS f. The suspension is mixed thoroughly
to the WPRBC to prepare the to produce a homogenous solution.
desired concentration (in %) of the Check for the presence of clot in the
RCS suspension
g. Label the RCS according to the concentration and blood (antigen) type
METHOD B
a. Preparation of washed packed red blood cell
(1) Using a Pasteur pipette, transfer 2 mL (2) Fill the tube half- full with NSS
of blood into a plain test tube
(3) Cover the tube with parafilm and mix (4) Centrifuge for 1 min. at 3400 rpm
by gentle inversion
(5) Aspirate the supernatant using Pasteur (6) Repeat steps 2 →3→4→5 three times (washing
pipette. This can also be done by discarding phase)
the supernatant as quickly as possible in (7) After the last washing , completely aspirate the
order not to disturb the cell button supernatant using as pasteur pipette
b. Preparation of tube with fixed volume of fluid
(1) Using a 0.1 mL serological pipette, (2) Deliver exactly 0.1 mL into a
aspirate NSS up to the highest plain test tube (use plain tubes
mark the calibrate the volume at with identical size for the whole
0.1 mark procedure
(3) Mark the level. The volume in this tube shall be used for comparing drops
administered that will be equivalent to 0.1 mL
c. Preparation of RCS for various concentration
(1) Label 4 plain test tubes (with identical size ) as 2% RCS , 3% RCS, 4% RCS and 5% RCS
(2) Using only one Pasteur pipette, deliver drops of the washed packed red blood cell to the
labeled tubes until it reaches a volume equal to 0.1 mL by comparing it to the volume of
the tube prepared in step B (Preparation of tube with fixed volume of fluid )
(3) Using 2.0 mL or 5.0 mL serological pipette, deliver exactly the following required
volume of NSS to the labeled tubes containing 0.1 mL of washed packed red blood cells
2% RCS → add 4.9 mL NSS
3% RCS → add 3.2 mL NSS
4% RCS → add 2.4 mL NSS
5% RCS → add 1.9 mL NSS
(4) Cover the tubes with parafilm then mix gently. These are the red cell suspensions at
different concentrations
NOTE : A proper red cell suspension has a tomato red color
