A typical procedure to obtain
mesophyll protoplasts:

  • Healthy leaves, removed from the plant and washed, sterilized and rewashed in sterile distilled water (subsequent procedures are conducted under aseptic conditions).
  • When leaves are in the final rinse, lower epidermis is peeled from the leaves or the lower epidermis is scored several times.
  • Cut the leaves into small sections, and transfer to filter sterilized enzyme solution.
  • Seal the dishes wrap them with aluminum foil (leave overnight).
  • Teased gently with forceps to release the protoplasts.

    • Purify protoplasts (filtration, centrifugation, and washing)

Syringe used to filter sterilize enzyme solution