the Ascocarp in Monascus. 169 
removed by treatment with hydrogen peroxide ; and it was 
then stained with safranin for twenty-four hours. After de- 
hydration with absolute alcohol, the stained material was 
placed in xylol for a few minutes until perfectly transparent, 
then teased out carefully and mounted in xylol-balsam. 
The reason for cultivating the fungus in a thin layer of 
liquid in a Petri dish was that, when it was grown in any 
considerable depth of liquid, growth took place more slowly 
and ascocarps were produced more irregularly and less 
generally. 
The material used for section cutting was obtained from 
plate-cultures on beer-wort agar. Such cultures showed the 
preliminary stages of fructifications after forty-eight to 
seventy-two hours from infection with conidia at 25 ~ 27 °C. 
At intervals of twelve hours portions of the agar, containing 
the fungus were fixed, washed, and hardened as above. After 
passing successively through half-alcohol and half-xylol, pure 
xylol, and xylol-paraffin into paraffin, series of sections were 
cut by microtome. These were treated in the usual manner, 
decolorized by hydrogen peroxide and stained by the Flem- 
ming triple stain. In a few cases the iron-alum haema- 
toxylin stain was used, but apart from bringing out the 
conspicuous nucleoli it was unsatisfactory. The cultures on 
beer-wort agar have a considerable advantage over those in 
liquid media, in that the formation of ascocarps is general 
and plentiful throughout, and at the same time at any given 
moment the majority of them are at approximately the 
same stage of development. Control is therefore easy. They 
are, however, unsuited to the previous method of examination, 
since satisfactory teasing of the agar material is impossible, 
the agar itself at the same time partially obscuring the 
fungus. 
The Development of the Ascocarp. 
The results obtained from the examination of living material 
will first be considered. 
In- a hanging drop-culture of beer-wort agar, infected with 
