Radiopharmaceuticals

	7: 111In LABELLING OF PLATELETS (using 111In-tropolonate)      (Merseyside And Cheshire Radiopharmacy Services)
	Materials Required
	111In-Indium Chloride solution.	1 x sterile end piece
	1 x 50ml syringe	2 x 60ml tubes
	1 x 10ml ACD Solution	2 x 30ml tubes
	1 x Tropolone in Hepes amp.	2 x 1ml syringes
	3 x kwills	3 x 10ml syringes
	1 x 21g needle	2 x 5ml syringes
	Method
1.	Obtain 50-60ml whole blood anticoagulated with ACD solution (Acid Citrate Dextrose) 1.5 parts ACD to 8.5 parts Blood.
2.	Distribute the blood between two sterile 60ml tubes and centrifuge at 90-100g ( 1000 rpm) for 10 minutes.
3.	Remove the supernatant taking care to avoid disturbing the red cell layer, and transfer to a 30ml Sterilin Universal tube.
4.	Adjust pH of supernatant to 6.5 by adding ACD solution. Check pH with narrow range pH paper.
5.	Centrifuge supernatant at 640g ( 2000rpm) for 10 minutes.
6.	Carefully remove the supernatant (without disturbing the cell pellet) and transfer to a second Sterilin tube. Label as Platelet Free Plasma PFP. and set aside.
7.	Resuspend the cell pellet with 0.3ml PFP.
8.	Add 50 microlitres of Tropolone solution followed immediately by 10-15MBq of 111In-indium Chloride.
9.	Incubate at room temperature for 5 minutes.
10.	Add approx. 5ml of PFP, mix then centrifuge again at 640g (2000rpm) for 10 minutes.
11.	Remove the supernatant and set aside for calculating the labelling efficiency.
12.	Carefully resuspend the cell pellet with approx. 5ml PFP, draw up into the syringe and cap with a sterile end piece.
13.	Calibrate activity in the syringe, label and calculate labelling efficiency.
	---------------------- For radiolabelling platelets with 111In-oxine refer to 111In-oxine Package Insert (Amersham Healthcare)

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