Culture of single Cells

(Bergmann’s cell plating technique)
In this technique, free cell are suspended in a liquid medium (if cell aggregates are there, the culture is filtered), and a culture medium with agar (0.6-1%) is cooled and maintained at 35ºC in a water bath. Equal volumes of these liquid and agar media are mixed and rapidly spread in a Petri dish, so that cells are evenly distributed in a thin layer, after solidification. The Petri dishes are sealed with parafilm and examined with inverted microscope to mark single cells (marking is done on outer surface of the dish). The plates are incubated in dark at 25ºC and cell colonies developing from marked single class, are used to obtain methods (e.g. filter paper raft-nurse technique; microchamber technique, etc.) have also been developed to grow individual cells.