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[Radial complement fixation test and convex lens method for the determination of antibodies in tissues]
The radial complement fixation test has to be made in forms in 1.4% layer of agar in physiological saline in a Petri dish. These forms have to be of the same thickness as the reaction itself, usually about 1 mm. In the first layer, it is recommended to mix antigen with complement and sheep erythrocytes in agarose and this mixture is to be poured over the tissue fragments examined. Following the primary incubation (at room temperature overnight), another mixture of haemolysin in agarose is poured over the first layer in form of optical convex lenses. The haemolytic phase is performed at room temperature, too. The complement is titrated together with the optimal amount of antigen and a drop of positive serum is placed in the well just in the centre of the first layer. In this way an optimal dilution of complement is detected, which is capable of producing a full hemolysis outside the zone of the positive serum diffused during the first, primary incubation period, whereas the zone itself remains untouched with the haemolytic activity of complement.
The radial complement fixation test has to be made in forms in 1.4% layer of agar in physiological saline in a Petri dish. These forms have to be of the same thickness as the reaction itself, usually about 1 mm. In the first layer, it is recommended to mix antigen with complement and sheep erythrocytes in agarose and this mixture is to be poured over the tissue fragments examined. Following the primary incubation (at room temperature overnight), another mixture of haemolysin in agarose is poured over the first layer in form of optical convex lenses. The haemolytic phase is performed at room temperature, too. The complement is titrated together with the optimal amount of antigen and a drop of positive serum is placed in the well just in the centre of the first layer. In this way an optimal dilution of complement is detected, which is capable of producing a full hemolysis outside the zone of the positive serum diffused during the first, primary incubation period, whereas the zone itself remains untouched with the haemolytic activity of complement.