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

Takashi Murata

Introduction

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 the glass surface of a glass-bottomed dish (IWAKI 3910-039). The thickness of the agar medium is minimized to obtain good optical conditions.　 Additionally, having a large mass of agar medium connected with the thin layer of the medium, in which the protonemata grow, allows continuous nourishment and growth of the gametophyte.

Methods

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

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