VELOSO, MARY RAFFAELE G BSMT 3D TABLE 5-MA’AM BANA-AY 02/11/21

EXERCISE 2: PREPARATION OF RBC SUSPENSION

PART I: WASHING OF RED CELLS (50PTS)

ILLUSTRATION PROCEDURE
Step 1: Label a 10ml test tube with “2% RBC Suspension” and/or
“5% RBC Suspension” and put it aside.

Step 2: Once blood collection is done, centrifuge your EDTA

tube to separate the plasma. Aspirate the plasma with a
Pasteur Pipette being careful not to disturbed the packed RBC
in the bottom. Transfer the plasma on a separate container
with 10% Sodium Hypochlorite.

Step 3: When most of the plasma has been removed from your
EDTA Tube, forcefully add NSS from a wash bottle up until the
liquid level is ½” from the rim.

Note: This is done without placing the tip of the wash bottle
into the tube to avoid contamination.

Step 4: Cover the top of your EDTA tube with its corresponding
tube top, invert, and mix.

Note: Make sure that all RBCs packed at the bottom are
dislodged until a homogenous mixture is formed.
Step 5: Centrifuge your EDTA tube with a balance tube for 1
min at 3400 RPM.

Note: Do not open the lid until the centrifuge has stopped
spinning.

Step 6: Using a transfer pipette (Pasteur Pipette), carefully

remove the supernatant fluid and discard it into a large
beaker with disinfectant (10% Sodium Hypochlorite)

Note:
• If supernatant fluid is still pinkish, the wash phase is
not yet complete.
• Be careful not to disturb the packed RBCs in the
bottom since we want to preserve them as much as
possible.

Step 7: Repeat steps 2-6 two more times, making a total of

three washes. Once done, label your EDTA tube with
“Washed RBC” as washed RBC cell is no different from
packed red cells with plasma.
PART II: DILUTION

1. Show your computations as to how to prepare

a. 2% Red Cell Suspension
i. Given: Conc.: 2% Red Cell Suspension, Total Volume Needed: 7ml, 100ml of NSS
ii. Calculations:

iii. We need 0.14ml of washed RBC and 6.86ml of NSS from the stock 100ml of NSS to make a 7ml of 2%
Red Cell Suspension.

b. 5% Red Cell Suspension

i. Given: Conc.: 5% Red Cell Suspension, Total Volume Needed: 7ml, 100ml of NSS
ii. Calculations:

i. We need 0.35ml of washed RBC and 6.65ml of NSS from the stock 100ml NSS to make a 7ml of 5%
Red Cell Suspension.

2. Write a short procedure as to prepare the 2% Red Cell Suspension.

a. Step 1: Prepare your separate tube labelled “2% RBC Suspension” from the previous procedure.
b. Step 2: Aspirate 0.14ml of Washed RBC from your EDTA tube and transfer it to your 2% RBC suspension tube
using a 1ml pipette.
c. Step 3: Aspirate 6.86ml of NSS and transfer it to your 2% RBC suspension tube.
d. Step 4: Mix the RBC suspension by inversion
e. Step 5: Once done, the end product of your 2% RBC suspension should yield a homogenous cherry red color.