282 RUTH STOCKING LYNCH 



plant, or from one explant when more than one was present, 

 liver cells only growing out from another part of the same explant 

 or from another explant. Wandering cells occurred anywhere 

 in the cultures, on or among the other cells or entirely isolated 

 from them. They were much more abundant in cultures show- 

 ing little or no growth than in those actively growing. 



According to Beck^ clasmatocytes are rare in the liver of a 

 seven-day chick embryo, and the wandering cells which appeared 

 in our cultures seem to differ somewhat from them. 



METHOD 



Explants were made from embryos of five to eighteen days' 

 incubation. These were examined at all stages of cultivation 

 from the time of planting to the death of the culture. The 

 culture medium used, Locke-Lewis solution (Locke's solution 

 plus 0.5 per cent dextrose plus 10 to 20 per cent chicken bouillon), 

 was the same throughout the entire series of experiments. 



The living cultures were studied with and without the use of 

 the various vital dyes. Janus green, neutral red, and trypan 

 blue were used. Cultures at various stages of growth were 

 fixed in osmic acid vapor, in Zenker without acetic acid, in Zenker- 

 formol, in Schaudinn, Bouin, and in iodine vapor. The first 

 two fixatives were the most successful. Preparations were 

 stained in hemateine and hematoxylin alone and in combination 

 with eosin, carmine and methyl green; in carmine, iodine, 

 crystal violet and Gram's stain; in safranin and licht griin, and 

 in acid fuchsin and methyl green. The results obtained with 

 carmine, iodine or crystal violet were unsatisfactory. Neither 

 of the two glycogen stains, carmine and iodine, gave definite 

 staining of the granules; with each of them the nucleus, nucleolus, 

 and debris stained deeply and the cytoplasm lightly. 



All drawings were made with the camera lucida, from the 

 living cell. The photographs are of the fixed preparations. 



2 Beck, C. S. 1919 The relative distribution of clasmatocytes in the various 

 organs of the seven-day chick embryo. Anat. Rec, vol. 16, p. 143. 



