Culture of Isolated Single Cells - Free cells isolated either from plant organs (mesophyll tissue) or cell suspensions are grown as single cells under in vitro conditions using a suitable medium. This process, called plating, is of particular importance when attempting to obtain single cell clones.

The plating technique developed by Bergmann (1960), (Razdan, 1993) is the most popular one for plating of single cells. Free cells are suspended in liquid medium at a density twice the finally desired plating cell density.

Equal volumes of the media containing single cells and a melted (30-35°C) agar medium (0.6-1%, w/v) are mixed and rapidly spread out in petri dishes in such a manner that the cells become fixed in an evenly distributed thin layer (1 mm thick) after the agar has cooled and solidified.