| Popis: |
Publisher Summary This chapter discusses plaquing and recovery of individual antibody-producing cells. The method that involves plating of antibody-secreting cells in methyl cellulose permits efficient recovery of the individual plaque-forming cells and their subsequent growth into large cultures. Usually plaque formation is done in agar or agarose between glass slides or in carboxymethyl cellulose. Under such conditions, it is difficult to recover and grow the individual plaque-forming cells. In this method, the red blood cells are plated in agarose, thus providing a firm support in which the red cells cannot diffuse and aggregate. On top of the red cell–agar mixture are plated the plaque-forming cells in methyl cellulose. After plaque formation, the individual plaque-forming cell is transferred with a micropipette to ordinary liquid medium. Plaques are perceived under different conditions at about the same efficiency, although at high cell concentrations, the non-plaque-forming cells impart a turbid background and the particular plaque-forming cell cannot be recognized. |
