c. One-Tube Crossmatch Procedure.
STEP 1. Place two drops of recipient serum in a labeled tube.
STEP 2. Add one drop of a 2 to 5 percent saline suspension of donor
(3) STEP 3. Centrifuge; examine for hemolysis, agglutination; grade and
Cold agglutinins may cause difficulty with this procedure. The two-tube
crossmatch may resolve the difficulty.
(4) STEP 4. Add 2 to 3 drops of 22 to 30 percent albumin according to
manufacturer's directions; mix. Centrifuge, incubate 15 to 60 minutes at 37C, read and
record the results.
(5) STEP 5. Centrifuge immediately upon removing from the incubator;
examine for hemolysis, agglutination; grade and record results.
(6) STEP 6. Wash three or four times with saline. After last wash, decant
completely. Add 1 to 2 drops of antiglobulin serum and mix.
(7) STEP 7. Centrifuge, examine for agglutination, grade and record the
results. (Use of optical aid is optional at this step.)
(8) STEP 8. Add one drop of known sensitized cells to all negative tests.
Centrifuge, examine for agglutination, and record result. If no agglutination is seen, the
antiglobulin phase must be repeated. If no hemolysis or agglutination is seen in any
phase, the blood is considered compatible.
d. Two-Tube Crossmatch Procedure.
STEP 1. Place two drops of recipient serum in each of two labeled
(2) STEP 2. Add one drop of a 2 to 5 percent saline suspension of donor
cells to each tube, and mix. Centrifuge, read, and record results.
(a) Incubate one tube 15 to 30 minutes at room temperature.
(b) Centrifuge; examine for hemolysis, agglutination; read and record