Enzyme treatment for digestion of cell wall. Protoplasts were isolated as follows using a separating mesh, because any mechanical stresses such as shaking, filtering, and even gently swirling in ordinary isolation protocols should be avoided. . B. 1 hr after incubation with the enzyme solution. C. 2 hr after incubation with the enzyme solution. These freshly isolated protoplasts often have irregular shapes, and the cytoplasmic streaming has generally ceased.
Next fig shows several isolated giant protoplasts above.
Generally, after 10 to 30 min protoplasts recover their active cytoplasmic streaming and become reasonably good spheres. Next figure shows protoplasts isolated from carrot root cells by a similar method as Nitella . These are also large enough and active enough for electrophysiology. Insertion of electrodes and Results . More results on membrane potentials 4.