236 BOTANICAL GAZETTE locroser 
G from 5-25 sec.; a longer time may do no harm but probably 
10-15 sec. is always sufficient. Wipe away excess of liquid and 
flood the slide twice with absolute alcohol, allowing the second 
lot of alcohol to remain on the slide until sufficient of the gentian- 
violet has been removed. The time required will depend on the 
material, the length of time it was in the gentian-violet, and the 
result desired. Drain rapidly with filter paper and flood with — 
clove oil for one minute; drain, follow by cedar oil and cover — 
in xylol balsam. If properly stained the host cell wall should 
be a light violet, the chromatin of the spirem and the chrome 
somes blue, nucleolus and centrosomes red, and cytoplasm 
slightly yellowish. 
Hematoxylin stain was used, applied after the method of 
Heidenhain (iron-alum 2 hrs., hematoxylin 12-18 hrs., followed 
by the slow extraction of iron alum till the proper degree 1s 
reached). This treatment gave some beautiful results in contrast 
with the Flemming stain, and was particularly valuable in demon 
strating the achromatic portion of the nuclear figure. Hartog’s 
nigrosin-carmine stain, as used by Wager and Berlese, ¥# 
tried repeatedly on corrosive sublimate material, but the results 
were far inferior to those afforded by Flemming’s tpl = 
Heidenhain’s hematoxylin stains. However, it is possible © ” 
demonstrate, even with this combination, the presence of a - 
nuclei in the oosphere and in the antheridial tube, and to rec 
soni ee ; : ‘c figures. thet 
ognize the principal features of the mitotic Mg! - Ehrlich, 
stains employed were Delafeld’s haematoxylin, Biondi’ — 
: = inferior in thet 
and cyanin-erythrosin, but they were distinctly 1n 
results. 
The following stain reactions were presente‘ - 
arations, and were attained by the Flem 
unless otherwise stated: chromatin blue or v!0 
Heidenhain’s hematoxylin ; nucleoli red, black wis pe datk 
hematoxylin; centrosomes as nucleoli; ie n page 16} 
: : . e Oo if ey 
blue ; cytoplasm yellowish ; granules, mention ee EE 
are only seen in preparations stained with Heide 
toxylin, and then black. 
