Sterile technique must be used throughout. It is important that all equipment used for the preparation of reagents be thoroughly cleaned to assure the absence of trace metal impurities. The user should wear waterproof gloves during the handling and administration procedure

  1. Withdraw from the patient 30-50 ml blood [preferably fifty (50) ml] using aseptic venipuncture technique using the 60 ml syringe fitted with a 19-gauge or 20-gauge needle and containing approximately 1000-1500 units heparin in 1-2 ml. Blood withdrawal should be smooth and slow so as not to produce bubbles or foaming
  2. Remove and dispose of the needle and replace with syringe cap. Gently mix the contents of the syringe and label with the patient’s ID, date and time.
  3. Upon receipt of the full syringe for processing, the contents should again be gently mixed.
  4. Clamp the syringe barrel to the ring stand in an upright (needle side up) position and tilt the syringe 10-20 degrees from its position perpendicular to the bench.
  5. Allow the red cells to sediment 30-60 minutes, depending upon when the supernatant [Leukocyte Rich Plasma (LRP)] looks clear of red cells.
  6. Replace the syringe cap with an infusion set
  7. Collect the plasma (LRP) in the centrifuge tube marked “WBC” by expressing the LRP through the catheter tubing making sure not to get any red cells into the WBC tube
  8. Immediately centrifuge the capped WBC tube at 400-450 g for 5 minutes
  9. Transfer the supernatant to the leukocyte poor plasma “LPP” tube leaving behind 0.5-1.0 ml supernatant to cover the white cell button. (Note: the button often contains a small number of red cells and may appear red.)
  10. Wash the white cell button with 4-6 ml Sodium Chloride (0.9%) Injection USP. Resuspend the cells by gentle swirling.
  11. Centrifuge the capped “WBC” tube at 400-450 g for 5 minutes (alternatively, 150 g for 8 minutes) and discard all but 0.5-1.0 ml of the supernatant to cover the cells.
  12. Add 5.0 ml Sodium Chloride (0.9%) Injection USP. Resuspend cells by gentle swirling.
  13. With the shielded syringe, draw up approximately 22.2 MBq (600 μCi) Indium In-111 Oxyquinoline. Check the amount of radioactivity in a dose calibrator set for Indium In-111 and record for labeling efficiency calculations. Parenteral drug preparations should be inspected visually for particulate matter and discoloration before administration.
  14. In several additions, add the Indium In-111 Oxyquinoline to the “WBC” tube, gently swirling after each addition.
  15. Set the lab timer for 15 minutes and allow the capped “WBC” tube to incubate. Swirl the cell preparation several times during the incubation.
  16. With a sterile plastic syringe, add half of the saved LPP (or about 8 ml) from the LPP tube. Cap and gently swirl the contents of “WBC” tube to resuspend the cells
  17. Centrifuge the “WBC” tube at 450 g for 5 minutes (or 150 g for 8 minutes). Decant supernatant into the “Wash” tube leaving behind about 0.5 ml of the supernate to cover the cells.
  18. Assay the activity in the “WBC” tube and in the “Wash” tube in a dose calibrator and record.
  19. With a sterile plastic syringe add the remaining LPP to the cell button and gently resuspend by swirling. With a sterile syringe fitted with a 19-gauge needle, resuspend the cells by drawing the cells up into the syringe and expressing the suspension against the tube gently one or twice. Alternatively, draw up the cells into a syringe fitted with the filtered 19-gauge needle, and replace the needle with an unfiltered 19- or 20-gauge needle.
  20. . Reserve in the “WBC” tube a minimum amount of white cell suspension for a WBC count. A microscopic examination should also be completed to observe for clumping. Draw up the patient’s dose 7.4 to 18.5 MBq (200-500 μCi) and check the syringe in the dose calibrator. Record the measurement