What is the Function of TwinSpin in Buffy Coat Preparation?

EL CAJON, Calif. - Feb. 24, 2023 - PRLog -- 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

TwinSpin centrifuge tubes with PBMC already inside Peripheral blood mononuclear cells (PBMC) can be isolated from bone marrow and whole blood using Spin Medium. The TwinSpin is made up of an inner tube and a standard 15. The inner tube's open bottom is buried in the density gradient medium (DGM). The blood sampling tube can be directly pipetted into the TwinSpin with anticoagulated blood or bone marrow.

The sample is placed on top of the DGM inside the inner tube. Leukocytes, lymphocytes, and PBMCs are separated from unwanted erythrocytes and granulocytes during density gradient centrifugation, depending on the density gradient used (Leuko Spin, PBMC Spin, PBMC 24+, or PLT Spin Medium).

Pre-filled Density Gradient Medium

1. Leuko Spin is used to separate leukocytes from bone marrow or whole blood.
2. PBMC Spin: Used to separate PBMC from bone marrow or whole blood.
3. PBMC 24+ Spin - For the separation of PBMC from bone marrow or whole blood older than 12 hours.
4. PLT Spin is used to separate platelets from bone marrow or whole blood.

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.

1. Preparing a Buffy Coat fraction in your lab using fresh whole blood
2. Another option is to create the buffy coat on your own. So just adhere to this quick protocol:
3. Combine one part washing buffer with one part whole blood.
4. Using the brake off, centrifuge the diluted whole blood for 10 minutes at 200 x g.
5. Remove the interphase leukocytes (buffy coat)

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