(1) Heparinized blood in a tube is left stand for about 1 our.
(2) The leukocyte-rich upper layer is taken out and splead over a polycarbonate film, which placed on the milipore filter pre-soaked with autologous plasma.
(3) Leave for about 1 our to allow the cells to sendiment.
(4) Remove plasma by the use of Pasture pipet or microtip.
(5) Cover with polycarbonate film (5 micrometer thick).
(6) Confirm the cells in adequately monolayed under the discecting microscope. If cells are too thick or non-homogeneous or clumped, splead evenly by glas rod and tilt for a while so that excess plasma and cells are to flow out. Tehn, remove the flowed out plsma. However, when the preparation is kept slant for further about 1 hour in a moisture chamber, nearly monolayered cell spread is usually obtained.