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