1 68 Barker . — The Morphology and Development of 
Methods. 
The methods used for determining the development and 
structure of the ascocarp in this fungus were twofold, viz. 
{a) The direct observation of the developing fructification 
under a Zeiss dz °il immersion lens (eye pieces 4 and 8) in 
hanging drop cultures. The hanging drops were made of 
beer-wort agar (beer-wort 98 per cent., agar 2 per cent.), and 
were infected with conidia from pure cultures of the fungus. 
The temperature was kept constant at 27° C., this degree 
being much more suitable for the rapid development of the 
ascocarps than the ordinary room temperature, growth prac- 
tically ceasing under i8°C. 
The results of these observations were checked by the 
examination of living material, containing developing asco- 
carps in all stages, under similar powers of magnification. 
In this manner the results of observations of comparatively 
few ascocarps, the growth of which had been watched from 
the start to the formation of the ripe spores, were generalized. 
(b) The examination of material after killing and staining. 
The fixing fluid used was the weak Flemming mixture. 
Ascocarps of various ages were not only examined whole, but 
also in series of sections, cut by the microtome. The material 
for these two methods was obtained by different means. 
For the examination of the entire ascocarps cultures of the 
fungus in a pure condition were made by infecting a tube of 
beer-wort with conidia, pouring the infected wort into a sterile 
Petri dish until the liquid covered the bottom of the dish to 
a depth of one-eighth to one-quarter of an inch, and then 
allowing growth to take place at a temperature of 25-27 0 C. 
The first stages of the ascocarps made their appearance after 
forty-eight hours ; and at the end of three and four days 
material was picked out with a sterile platinum needle and 
fixed. Such material contained ascocarps in all stages. 
After fixation, it was washed for twenty-four hours in running 
water and then hardened in a series of alcohols ; the blacken- 
ing produced by the osmic acid of the fixing fluid was 
