CROSSMATCHING

(Major and Minor)

- establish compatibility or incompatibility between the donor and the recipient’s blood. Patient is the recipient.

1. MAJOR CROSSMATCH  LABELLED AS TUBE 1

2 drops PATIENT’S SERUM (collect blood from the patient/donor using red top or EDTA tube
+
1 drop DONOR’S RBC SUSPENSION (exact or estimated) if estimated, 1 drop of RBC + 19 drops of NSS
 PS2DR1

2. MINOR CROSSMATCH  LABELLED AS TUBE 2

2 drops of DONOR’S SERUM
+
1 drop PATIENT’S RBC SUSPENSION
 DS2PR1

THIS IS ALREADY CONSIDERED AS THE FIRST PHASE OF CROSSMATCHING (SALINE PHASE)

------------------------------ EXAMINE MACROSCOPICALLY -------------------------------
FOR AGGLUTINATION OR HEMOLYSIS  RECORD RESULTS

Add 2-3 drops of Bovine Albumin to both tubes (colorless) / LISS (Low ionic strength saline) to both tubes
incubate = 15 -30 minutes (basi diri pa lang ma form ang agglutination), the longer the incubation, the better but
should not exceed 30 minutes because it will cause evaporation of the specimen. If no incubator, water bath
may be used.
Centrifuge = 15 seconds, take out tubes and resuspend.
THIS IS THE SECOND PHASE OF CROSSMATCHING (PROTEIN PHASE)
------------------------------ EXAMINE MACROSCOPICALLY -------------------------------
FOR AGGLUTINATION  RECORD RESULTS
Washing =3X with NSS (about ¾ full of the tube) = Centrifuge (5 mins)
NSS is an isotonic solution. If hypertonic (crenated) and hypotonic (swollen) changes the shape of the cells.
1st wash = reddish, then decant
2nd wash = slightly reddish, then decant
3rd wash = clear, then decant
THIS IS CONSIDERED AS THE PREPARATION FOR THE THIRD PHASE
--------------------------------------------------------------------------------------------------------
Add 1 drop of AHG (green) to both tubes – bridge the immune complexes that has been formed. IgG is
incompetent antibody to form agglutination unless AHG is added that will bridge the gap between the two IgG’s.
Centrifuge = 15 seconds
--------------------------------EXAMINE MACROSCOPICALLY = Hemolysis
MICROSCOPICALLY = Agglutination

compatible = NO AGGLUTINATION/HEMOLYSIS
incompatible = WITH AGGLUTION / HEMOLYSIS
THIS IS THE THIRD PHASE OF CROSSMATCHING (ANTIGLOBULIN PHASE)