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 gentl swirling in ordinary isolation protocols should be avoided. . B. 1 hr after the incubation with the enzyme solution. C. 2 hr after the incubation with the enzyme solution. These freshly isolated protoplasts are often not good sphere, and the cytoplasmic streaming is generally still stopping.
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.