Mechanical Method of Isolation of Protoplast - In mechanical method, cells are kept in a suitable plasmolyticum (in plasmolysed cells, protoplasts shrink away from cell wall) and cut with a fine knife, so that protoplasts are released from cells cut through the cell wall, when the tissue is again deplasmolysed.

This method is suitable for isolation of protoplasts from vacuolated cells (e.g. onion bulbs, scales, radish roots). However, this method gives poor yield of protoplasts and is not suitable for isolating protoplst from meristematic and less vacuolated cells. The mechanical method, though, was used as early as 1892, is now only rarely used for isolation of protoplasts.