Fibronectin solution preparation

Fig. 3. Preparing a coverslip for live cell microscopy, (a) Dissolve 1 mg of bovine fibronectin In sterile water. After 1 h, add 4 ml of PBS and store 200 pg/ml fibronectin solution at 4°C. (b) Remove gaskets trom plastic permanox 8-well chamber. Cut epoxy mold squares and stick to No. 1.5 gold seal cover glass. Add 125 pi ot tibronectin, let sit for 1 h, rinse once with RPMI culture medium, and store In RPMI medium until ready to Image.

Red blood cells, erythrocytes, were used because of their low and reversible adhesion. Cells were prepared from three species, human blood from North Staffordshire Hospital, fresh horse blood in EDTA, and fresh rat blood from Central Animal Pathology Ltd. Each blood sample was washed six to seven times in phosphate buffered saline to remove the nonred-cell components, before suspending in physiological saline solution, then examined by both optical and Coulter tests. Each species of cell was treated in three ways to judge the effect of surface adhesion molecules by adding glutaraldehyde, fibronectin, and papain. 

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