Isolation of Single Cells - Single cells can be isolated from plant organs, particularly leaf, either by mechanical or enzymatic means or they can be separated from suspension cultures.

For mechanical isolation, leaves are cut into small (1 cm2) pieces, ground in a suitable culture medium using pestle and mortar, the resulting homogenate is filtered through muslin cloth, and the cells are pelletted out by centrifugation.

In the enzymatic method, the lower epidermis of leaves is peeled off and the leaves are cut into moderate pieces (4 cm2), which are incubated in a macerozyme (0.5%) or pectinase solution. Partial vacuum may be used to facilitate the entry of enzyme into the tissue, and medium solution may be changed during the treatment.