Flow sorting and culture of protoplasts: Conditions for high-frequency recovery, growth and morphogenesis from sorted protoplasts of suspension cultures of nicotiana

Plant Cell Rep. 1984 Aug;3(4):151-5. doi: 10.1007/BF00270211.


Protoplasts were prepared from suspension cultures of Nicotiana tabacum cv Wisconsin 38 that had been prelabeled with FITC. The protoplasts were subjected to flow sorting based on fluorescence content using a Coulter EPICS V Flow Cytometer - Cell Sorter. Conditions were established that allowed the recovery after sorting of approximately 30% of the initial protoplasts in a viable state. These were subsequently regenerated into calli that underwent shoot morphogenesis.