PRACTICUM 1

TITLE: PREPARATION OF RED BLOOD CELL SUSPENSION IN SALINE

INTRODUCTION
Saline solution have used in immunohaemotology lab for cell cell washing and to
prepare cell suspension. 1% sodium hypochlorite are prepared to clean up places
contaminated by blood. Suspensions of washed red cells having kwons antigens on their
surface are required for various tests in the laboratory. The washing of cells is necessary
to remove plasma, which may interfere with the reaction of the cells. In addition, blood
group substances in plasma may neutralise the antiserum leading to false negative
results. It is also necessary to use a red cells are suspended in isotonic solution of sodium
chloride, (0.15mol/L or 8.5g/L), pH adjusted to 7.0

OBJETIVE
1. Saline solution is used to wash cells and to prepare cell suspension
2. 1% sodium hypochlorite is used to clean up glassware and workbenches
contaminated by blood

MATERIAL
Human blood, Saline, Pasteur pipette, Centrifuge, Test tube, and Parafilm.

PROCEDURE:

A) Saline solution preparation (0.9% d/I)

1. Sodium chloride is weight 9.0g.
2. Sodium chloride is pour onto beaker
3. Distilled water is mixed up
4. The solution is pour into 1L to volumetric flask/
5. The beaker is washed with water and saline is poured in the saline into flask.
6. Water is added until 1L
7. The container are kept and labelled

B) PREPARATION OF 1% SODIUM HYPOCHLORITE FROM 10% PREPARED SOLUTION

1. 10ml of 10% sodium hypochlorite is prepared into 100ml sylinder
2. Water is added up until 100ml
3. Well mixed uo and bottle is kept in for storage
4. Container is kept and label.

C) PREPARATION OF CELL SUSPENSION

1. 0.2 to 0.5ml of blood is placed in a test tube.
2. The tube is filled up with saline until 1cm from the top.
3. The test tube is centrifuge for 1-2 minutes to obtain red packed cell.
4. The supernatant is removed by pouring off the saline in one quick, continuos
movement or by using a Pasteur pipette.
5. To resuspend the cells in the residual fluid, tap the tube. This constitutes one
wash
6. The procedure is repeated with the same cells at least twice. The last wash
showed clear, colourless supernatant with no signs of haemolysis.
7. Make 2% cell suspension, 1 drop of packed red cells and 49 drop of saline are
added.
8. Make 5% cell suspension, 1 drop of packed red cells and 19 drop of saline are
added.
9. Make 10% cell suspension, 1 drop of packed red cells and 9 drop of saline are
added.

RESULT
2% suspension 5%suspension 10%suspension
DISCUSSION
The purpose of this study was to validate a procedure for using red blood cell
suspensions stored for up to 10 days in an immediate spin crossmatch. Ten group A and
10 group B SAG-M donor unit segments were opened on day 0. The red cells were divided
into 2 tubes and each diluted with saline to a 2% to 5% cell suspension. The cell
suspensions were tested using a standard immediate spin technique against pooled
group A, group B, and group O plasma each day for 10 days. One set of cells was stored
refrigerated as a 3% to 5% cell suspension. Saline was removed from the parallel set of
cells before storage. All 480 tests that were expected to be negative were negative. On
2 separate occasions 1+ or 2+ reactions that disappeared before the cell button was
completely resuspended were noted. All 960 tests that were expected to be positive
were positive. All but 4 results were graded as 3+ or 4+. Four 2+ reactions occurred
with 2 units that were typed as A1 negative. Our results indicate that cells stored
refrigerated in normal saline as a 3% to 5% cell suspension may be used for immediate
spin crossmatch for up to 10 days.
It usually used for determine ABO blood grouping and for patient need blood
transfusion.

CONCLUSION
Based on the experiment conducted, student be able 2 saline solution is used to wash
cells and to prepare cell suspension. Besides that, student be able to understand and
use 1% sodium hypochlorite is used to clean up glassware and workbenches
contaminated by blood.

QUESTION
1. What is the importance of the study?
The purpose of this study was to validate a procedure for using red blood cell
suspensions stored for up to 10 days in an immediate spin crossmatch
2. What is Whartons jelly?
Wharton's jelly is a gelatinous substance within the umbilical cord, largely made up
of mucopolysaccharides hyaluronic acid and chondroitin sulfate. It also contains
some fibroblastsand macrophages. It is derived from Extra Embryonic Mesoderm.