On-Plant Method to Deliver Dye or Biomolecules Using VACNFs in a Rigid Substrate

To begin, remove the photo resist from the VACNFs by washing with 100%acetone for five minutes, followed by a five minute wash with 100%IPA, and then double distilled water for five minutes. Then, place the plant tissue to be impaled on a hard surface for support. Place a one microliter droplet of dye onto the surface of the plant tissue.

Place a VACNF chip with a rigid substrate down the top of the droplet, with the fibers oriented such that they will come into contact with the droplet. Using the flat side of a pair of tweezers, tap the chip, mark the area of the plant that the chip came into contact with using a soft tipped marker. Remove the VACNF chips after delivery.

Repeat these steps for controls, such as placing dye on the fiberless side of a film, using the film without dye, and using the fiberless side of a film. Store intact plants or excised plant organs in humid chambers in long day conditions. For excised organs, use a plastic Petri dish with wet paper towels.

Using this method, fluorescein dye was delivered to Arabidopsis leaves. Images taken at different time points, showed that the fluorescein dye was observed immediately after delivery. Similarly, delivery of a plasmid containing yellow fluorescent protein was achieved in Arabidopsis leaves.