1. Blood sample handling EHES Training Materials

2. Equipment Centrifuge Empty tubes for re- centrifugation (only needed if gel serum tubes are used) Storage tubes Pipette Storage boxes Disposable gloves

3. Before centrifugation of blood tubes After blood drawing store the tubes at room temperature in a vertical position at least 30 minutes for the blood in serum tubes to be clotted. Do not prolong the waiting time over 60 minutes! Adjust centrifuge speed 2000-2200 g RCF

4. Centrifugation of blood tubes (1/2) After the waiting period, centrifuge The serum tube The fluoride-citrate tube The 1 st EDTA tube Do not centrifuge the last two EDTA tubes designated for HbA1c and DNA extraction!

5. Centrifugation of blood tubes (2/2) Place the serum tube and the fluoride-citrate and the 1 st EDTA plasma tube in the centrifuge Check that all tubes are resting on the bottom of the centrifuge rack Check that the centrifuge rotor and tubes are in balance. Centrifuge the tubes 2000 g for 10 minutes at room temperature

6. Sample handling after centrifugation After the centrifugation, check that the serum is separated properly Pour the serum from the 2 serum collection tubes into the pooling tube, mix gently by swirling the contents Note! If one of the serum tubes is haemolysed, do not mix them together

7. Labelling storage tubes Place the storage tubes in a tube rack according to the pipeting scheme No need to label all tubes if blood collection is incomplete Place the bar code labels in an upright direction, so that the scale marks remain visible

8. Handling blood collection tubes with gel The temperature during centrifugation should be at least 20-22 °C Inspect the gel tube after centrifugation for: Gel surface horizontal Serum and cell layers separated clearly No red cells visible on top of the gel No fibrin strands in the serum Serum liquid, not clotted

9. Hemolysed samples Do not pool a hemolysed serum sample with non- hemolysed samples Several hemolysed samples can be pooled Document hemolysed aliquots Use the colour guide to document the degree of hemolysis

10. Pipette aliquots into storage tubes 2 x 1,5 ml serum from the pooling serum tube into the 3 ml plastic tube and 1,5 ml aliquots into the 1,5 ml storage cryotubes 2 x 1 ml from fluoride- citrate plasma tube into two 1,5 ml cryotubes 2 x 1,5 ml from the EDTA tube into two 1,5 ml cryotubes Close the cryotubes with caps and freeze the tubes at once

11. Transferring storage tubes into freezers Place the tubes without delay into the boxes in the freezer Label boxes before placing them into the freezer because labels will not stick on a wet/cold surface

12. Sample shipment To the national HES laboratory In dry ice Packed in leak proof secondary packaging To the EHES Reference Laboratory Packed according to the IATA regulations, according to Pecking Instruction 650 for diagnostic specimens Place one tube upside down in every box

13. Recording Record: The amount of the collected serum and plasma Visible attributes (haemolysis, lipemia, icterius) Deviations from the sample handling protocols, if any Your initials/personnel code to identify who handled the samples

14. Acknowledgements Slides Laura Lund, Päivikki Koponen Photographs Hanna Tolonen Demonstration Saara Vallivaara