Storyboard Text
- 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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