288 RUTH STOCKING LYNCH 



lines being" more or less lost in their crowding. In the flattened 

 cells the mitochondria appear separated and even somewhat 

 scattered, and are distributed throughout the endoplasm without 

 apparent plan, differing markedly in size and shape even within 

 the same cell. They all stained alike, however, with each stain, 

 just as in the living cell they all took up the janus green in a 

 uniform manner. 



Bile granules 



In the liver cells and wandering cells of these cultures there 

 sometimes occurred bright green masses of irregular form, usually 

 about the size of the nucleolus or a little larger. These were 

 probably bile granules. They were observed in cultures of two, 

 three, four, and five days' cultivation, from chick embryos of 

 seven, eight, and eleven days' incubation. The number of liver 

 cells showing them was always few; often only two or three in 

 an entire growth membrane. Wandering cells containing them 

 were much more numerous; in one culture made from an eleven 

 day embryo all the wandering cells showed large, bright green 

 masses on the second day of cultivation. It is probable that 

 these macrophagic cells took up these bile granules from the 

 broken down liver cells of the explant. 



The bile granules were evidently situated in the endoplasm 

 of the liver cells; they exhibited no movement and usually lay 

 close to the nucleus. They were never observed near the cell 

 periphery. No staining of these granules with trypan blue was 

 observed; in trypan blue cultures, cells containing both blue and 

 green granules were found. With neutral red, however, they 

 stained very deeply, more deeply than any other cell inclusion 

 observed. When a dilute solution of neutral red was added to an 

 unstained or a trypan blue culture the green granules changed to 

 orange, then to brown, to reddish-brown, and finally to a very 

 dark red. In cultures grown in a neutral red medium no green 

 granules were to be found. When neutral red cultures were 

 treated with potassium permanganate these masses gradually 

 lost their red color until after a time, varying from a half to one 

 and a half hours, they became a dark brown or even a brownish- 



