Peripheral blood tubes are immediately inverted 10 times and centrifuged at 4,200 x g, 7 min, with brake. The supernatant plasma is transferred to a new tube and centrifuged at 13,000 x g, 5 min with brake to pellet and remove the platelets. The supernatant platelet-free plasma is transferred to a new tube and centrifuged again at 13,000 x g, 5 min with brake to ensure there are no platelets left.
Hello
yes , it is possible . But can be isolated Platelet-free plasma (PFP) by a centrifugation using 1,500g for 20 minutes followed by a high speed centrifugation at 13,000g for 2 minutes (8) Because the centrifugation speed applied is crucial for the removal of platelets from plasma . Because PPP still contains some platelets, use of PFP may be preferred. However,it should be considered that some MPs might be lost during the high-speed centrifugation step.
During centrifugation the use of breaks should be avoided as much as possible and plasma should be carefully collected without disturbing the platelet layer to prevent remixing of the plasma with the platelets . MPs measurements should be performed preferably directly in fresh plasma to reduce MP loss and to preserve MP characteristics.
Platelets need to be removed from the plasma in order to avoid cellular activation
leading to inadvertent production of MPs. For this purpose, a proper centrifugal
speed should be applied for plasma preparation
For Plasma isolation
According to the Clinical and Laboratory Standards Institute (CLSI) .
plasma with a platelet count less than 10x109/L is obtained by centrifuging the blood at 1,500g for 15 minutes at room temperature (RT). To obtain platelet-poor plasma (PPP) CLSI recommends recentrifuging the plasma for another 10 minutes at 1,500g. We observed that PPPisolated by centrifuging blood once at 1,550g for 20 minutes still contained 2.8x109/L platelets. However, when the same blood sample was used to isolatePPP by performing a two-step centrifugation, each at 2,000g for 10 minutes, there were only 6.1x108 /L platelets present in the plasma.
Good Luck
The microparticle analysis by flow cytometry from blood does not require many centrifugation steps (unpublished work). With appropriate modification you can even work with whole blood staining protocol. Please let me know if you are looking for help in optimising your experiments on the same.
Sanjaya, can you provide details on analysis of microparticles from human serum
Sometimes several protocols for plasma isolation use the first centrifugation at RT and the second at 4°C. Does someone know why?
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