What is the Function of TwinSpin in Buffy Coat Preparation?
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A buffy coat is an accumulation of platelets and white blood cells (WBCs). We'll discuss how TwinSpin helps in the preparation of a buffy coat.
A sample of peripheral whole blood contains less than 1% white blood cells (WBCs) and platelets. When scientists spin the blood sample in a centrifuge, these WBCs and platelets combine to form their own layer that is suspended between the red blood cells (RBCs) and supernatant plasma. This thin layer is referred to as a buffy coat because of its color (yellowish to brownish).
We will also find out how TwinSpin density gradient centrifugation tubes are used in buffy coat extraction.
A Brief On TwinSpin Tubes
Spin Medium can be used to separate peripheral blood mononuclear cells (PBMC) from bone marrow and whole blood into TwinSpin centrifuge tubes. A standard 15-inch tire and an inner tube make up the TwinSpin. The open bottom of the inner tube is submerged in the density gradient medium (DGM).
Instructions for Using the TwinSpin Tube
1. Ensure that the (diluted blood) sample, the TwinSpin, and the centrifuge are all at room temperature.
1.1 Check that the inner tube is partially filled and immersed in the DGM. If not, maintain the vertical position while rotating the TwinSpin device.
1.2 Take out and throw away the transport stopper.
2. Include Sample Material
2.1 Pipette the sample material on top of the DGM in the inner tube by tilting the TwinSpin 45° and allowing the sample to run down the inside of the dropper.
2.2 Push the elastic cap firmly into the opening to close the TwinSpin. PluriSpin PLT Depletion can be used to reduce platelet contamination.
3. Spin
3.1 Centrifuge at 800 x g for 20 minutes at room temperature in a swing bucket rotor, brake off. If the blood is more than 4 hours old, centrifuge it for 30 minutes at 1000g.
4. Collect
4.1 Remove the inner cell separation tube by unscrewing it.
4.2 Push the elastic cup down to collect cells in the opaque layer in a new tube.
5 Wash (if necessary)
5.1 Add wash buffer to the reaction tube.
5.2 Cells are spun down for 10 minutes with 300 x g at 4 °C (no or small break).
5.3 Remove the supernatant and set the reaction tube on the table for 20 seconds.
Excess wash buffer will flow down the tube wall and accumulate at the bottom. Aspirate the majority of the liquid above the pellet. The liquid will appear foggy because it contains mostly platelets; aspiration will improve the washing result.
5.5 Carefully pipette the pellet with 1 ml of wash buffer.
5.6 Re-do steps 5.1–5.4.
5.7 Refill the pellet to the desired volume.
Preparation of buffy coat from scratch
Buffy coats can be prepared in two different ways. To separate the whole blood into red cells, plasma, and buffy coat, the whole blood donation is centrifuged on the one hand. The second option is to filter the blood and keep the leukocytes from circulating.
















