A detailed procedure for the intranuclear microinjection of plant protoplasts

Abstract

Procedures and equipment have been developed for the efficient intranuclear microinjection of plant protoplasts. The manipulation and culture of protoplasts was facilitated by the use of an environmentally controlled cabinet and video recording equipment. Microinjection rates of 80–100 nuclei/h were routine. The delivery of volumes as small as 10 −17 L was easily controlled with a custom-designed microinjection syringe system. Individual protoplasts were recovered with 90% efficiency with the assistance of microscopie grid patterns photolithographically applied to cover slips.