Question

Separation of most blood cells is difficult, if not impossible, to achieve because they have similar properties and/or densities. What procedure is used to separate T-cells of the immune system from the many other different types of white blood cells or spleen cells? What feature of the T-cell facilitates the isolation protocol? Describe how this procedure is performed.

Answer 1

Ficoll-paque density gradient separation is used to separate T cells from the rest of the cells. The procedure is performed by adding diluted whole blood in buffer like PBS or HBSS. It is then spinned over a Ficoll gradient, which results in majority of granulocytes to separate at the bottom of the tube along with red cells. The white blood cells remain in middle of the tube appearing as white ring also known as buffy coat. Plasma will form the top layer. The buffy coat can then be collected to obtain lymphocytes and monocytes. Finally a magnetic bead separation can be done followed by positive selections to carry out further isolation.