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  • Perform NOD and explain the procedure to the patient
  • 60 mL syringe containing 10 mL of ACD (acid citrate dextrose)
  • ACD (acid citrate dextrose)
  • Gain consent for a blood draw
  • Collect 50 mL of the patient’s blood, mix
  • Separate the blood/ACD into two samples
  • Transfer 15 mL of the blood/ACD from the original sample into a centrifuge tube
  • The rest remains in the syringe
  • 5-10 mL of Hetastarch (Hespan)
  • Hetastarch (Hespan)
  • Prepare to settle the RBC's
  • Add the hetastarch to the 45 mL of blood/ACD in the syringe
  • Mix the hetastarch with the blood/ACD in the syringe
  • Stand the syringe with the needle point up for 45-60 mins to let the RBC’s settle at the bottom of the syringe leukocyte rich plasma at the top (LRP)
  • Centrifuge the 15 mL hetstarch-free blood at high speed
  • Centrifuge
  • Remove and save the supernatant
  • The supernatant is called platelet-poor plasma (PPP)
  • Supernatant - platelet poor plasma (PPP)
  • Once the RBC’s have settled, transfer the LRP from the syringe into a centrifuge tube
  • Discard the syringe with RBC’s
  • Centrifuge the LRP at a slow speed
  • Centrifuge
  • Remove the leukocyte-poor plasma (LPP) from the top of the tube.
  • One tube of the WBC pellet and one tube of the leukocyte poor plasma (LPP)
  • Add 1ml of saline to the WBC pellet
  • Agitate the cells
  • Add sterile water to the WBCs
  • Agitate the cells for 20 seconds
  • Add 3-5% saline to the WBC’s to stop the lysis
  • Wash the RBC’s with normal saline
  • The mixture is centrifuged
  • Centrifuge
  • Remove the supernatant (RBC fragments and saline)
  • Resuspend the WBC’s again in 1 mL of saline
  • Leukocyte poor plasma (LPP)
  • WBC pellet
  • Add 1000 MBq of unstabilized 99mTc-HMPAO to the WBC’s
  • Saline
  • Incubate for 15-20 mins
  • Wash the cells by adding some of the hetastarch free PPP to remove excess 99mTc-HMPAO
  • Water
  • Centrifuge the cells
  • Remove the hot supernatant
  • 3-5%
  • Save the hot supernatant
  • Saline
  • Add 3-5 mL of the remaining PPP to the labelled cells to resuspend the cells
  • Saline
  • Centrifuge
  • Hot supernatant
  • Draw up the suspension in a 10 mL syringe
  • Measure the syringe of labelled cells in the dose calibrator
  • Dose Calibrator
  • Measure the tube of hot supernatant in the dose calibrator
  • Dose Calibrator
  • Calculate the labelling efficiency
  • Labelling efficiency
  • Have a second technologist identify the patient identify patient
  • Explain procedure and gain consent from patient
  • Re-inject the cells
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