2.7  Culture in a thin layer of agar medium spread in a glass-bottom dish

Takashi Murata




For continuous observation of protonemata at high resolution, we need to observe protonemata without damage under good optical conditions. For this purpose, protonemata are cultured along a glass surface of glass bottom dish. Thickness of agar medium is minimized to obtain good optical conditions.  In the other hand, large mass of ager medium connected with the thin layer of the medium, in which protonemata grow, allows continuous growth of gametophyte.



A small colony of protonemata is inoculated into a glass-bottom dish with medium, and then overlayed with a cellophane. After 3 or 4 days of culture, caulonemata began to develop.

Microscopy should be done from bottom.  Protonemata grow parallel to the bottom of the glass-bottom dish.