408 R. Ss. CUNNINGHAM 
a few scattered granules sometimes developed about the nucleus. 
In the cells covering the liver particularly and the other surfaces 
to some extent, the carmine granules, with increased staining, 
often developed into a long, oval belt which sometimes crossed 
the nucleus only on one side, and sometimes extended partially 
if not wholly around it. So that instead of having the cir- 
cumscribed perinuclear rosette which has been described as 
characteristic of trypan blue, these cells had a rosette in the 
form of a belt running all, or part way, around the nucleus antero- 
posteriorly. 
Finally, it seems important to emphasize the question of the 
orientation of the dye distribution in relation to the nucleus and 
the position of the cells in situ. These observations were made 
both upon living cells and upon sections, the latter being cut 
perpendicular to the surface. In sections the perinuclear rosette 
so typical of trypan blue was clearly seen as a patch of granules 
at either end of the nucleus, one or the other containing more 
blue corresponding to the area of localization. This was often 
extended somewhat over the surface of the end of the nucleus. 
But the characteristic arrangement was the principal amount 
of the blue close to one end of the nucleus. In the early staining 
with carmine the small sharply localized area of granules was 
usually between the nucleus and the surface of the cell, but 
placed laterally to the central axis of the nucleus, so that as 
the dye increased in amount with longer exposures the cross- 
sections showed red granules in both outer and inner zones of 
the cell. This again was the formation of the perinuclear belt, 
already described, from the lateral or anterolateral group of 
granules. Considering the relationship of the early areas of 
carmine staining to the center of the nucleus as distributed along 
its long axis, they were found to be usually somewhat nearer one 
end, but were often close to the center. As the carmine increased 
in amount to the maximum, the granules often covered the en- 
tire length of the nucleus, but more often formed the perinuclear 
belt. 
