288 LEO LOEB AND KENNETH C. BLANCHARD. 



(e) Trypanblue (1:5000). The cells appeared unstained. 

 The outgrowth was similar to control. 



(/) Janus green (1:4000). Appeared to be relatively toxic. 

 There was apparently a very slight diffuse green stain; neither 

 were granules distinctly stained nor were stained drops or 

 particles visible in the cells. Some amoeboid movement was 

 noticeable in this tissue. 



III. THE EFFECT OF ACID AND ALKALI ON THE DECOLORI- 

 ZATION OF AMCEBOCYTE TISSUE. 



Method. Amcebocyte tissue was prepared in stender dishes in 

 the way described previously. 2 The stain (dissolved in a n/2 

 NaCl solution) was poured over the tissue after the supernatant 

 serum had been poured off. Unless otherwise stated, the stains 

 were used in a dilution of 1 :5OOO. 



During the process of staining the tissue was kept in the ice 

 chest. The stained tissue was washed with n/2 NaCl solution. 

 Pieces of tissue were then cut out and placed in the small test 

 tubes which contained the solution, whose extractive power it was 

 desired to test. Before comparing the amount of stain given off 

 by the tissue in the various solutions, the pieces of tissue were 

 removed from the tubes and the solutions in the different test 

 tubes brought to the same hydrogen ion concentration and the 

 same volume. 



Neutral Red Tissue. Solutions of n/2OO, 72/500, n/iooo HC1 very 

 readily extract the neutral red from the tissue; the extraction 

 usually becomes noticeable to the naked eye within a period of 

 from ten to fifteen minutes or even somewhat earlier. In 

 solutions of n/2OO, n/iooo NaOH, not more than a trace of stain is 

 given off. Even after remaining in the alkaline solution for 48 

 hours in the ice chest very little stain was given off by the tissue. 

 In neutral solutions of n/2 NaCl likewise no or very little stain is 

 given off. Neutral red tissue which has been immersed in 

 n/iooo and in 72/500 HC1 for 48 hours, and has given off red stain 

 to the surrounding fluid, yields as much color again as it did the 

 first time, if transferred to a fresh acid solution. Other acids, 

 like n/iooo benzoic, butyric, lactic acid, extract the stain as well as 

 n/iooo HC1. 



