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Texto del Guión Gráfico
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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