TwinSpin centrifugation tubes pre-filled with Density Gradient Medium (DGM) can be used for an optimal separation peripheral blood mononuclear cells from whole blood and bone marrow. The TwinSpin is comprised of a standard 15 ml and an inner tube. The inner tube has an open bottom which is submersed in the DGM. Diluted anti-coagulated blood or bone marrow can simply be pipetted into the TwinSpin. The sample lays on top of the DGM inside the inner tube. During centrifugation, Leukocytes, lymphocytes and PBMCs are separated from unwanted erythrocytes and granulocytes, depending on the DGM. The result is an inter-phase enriched with target cells above the DGM. The erythrocytes will sediment through the density gradient media out of the inner tube at the bottom of the outer tube. When separation is complete, just remove the inner tube. The elastic cap functions as valve. By pushing the cap down, the collection tube becomes a pipette. The contents can be collected drop by drop. Prefilled with DGM of your choice.
Direction to fill the separation system:
- Unscrew the inner separation tube from the 15 ml tube, do NOT remove the transport stopper.
- Pipette 4.5 ml of a Density Separation Medium of your choice into the 15 ml tube.
- Insert the inner tube into the 15 ml tube – avoid pushing it down, it will slide in on its own.
- Screw airtightly the inner cup to the 15 ml tube.
- After inserting and screwing the inner tube make sure that there is Density Gradient Medium inside the inner tube. Perform a quick rotating movement, holding the tube in the upright
Directions for the use of the TwinSpin Tube
- Add Sample Material
Check that (diluted blood) sample, TwinSpin and centrifuge are all at room temperature.
Make sure that the inner tube is partially filled and submersed in the DGM
Remove and discard the transport stopper,Pipette the sample material on top of the DGM in the inner tube by holding theTwinSpin in a
inclined position at 45° angle and let the sample run downthe inside of the dropper. Close the TwinSpin with the elastic cap by pushing the cap firmly into the opening.
- Spin
Centrifuge for 20 minutes at 800 x g at room temperature within a swing bucketrotor, brake off. Using blood older than 4 hours centrifuge for 30 minutes at 1000g.
- Collect
Unscrew and remove the inner separation tube.Collect cells in the opaque layer in a fresh tube by pushing the elastic cup down.
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