SUPRARENAL GLAND—EFFECTS OF INANITION 223 
The suprarenals were removed immediately after the animals 
had been killed. The rats were killed by chloroform, excepting 
those used for the chromaffin tests (series F), which were 
killed by cutting their throats. One rat (F 6.2) died from acute _ 
inanition. 
In most cases the glands were fixed in Zenker’s fluid, embedded 
in paraffin, cut in serial sections (3 » to 5 uw), and stained with 
haematoxylin and eosin. Ina few cases other fixatives (formalin, 
Zenker-formol, Flemming’s) and other stains (iron-haematoxy- 
lin, Mallory’s anilin-blue connective-tissue stain, etc.) were em- 
ployed. In the series F and a few others one gland was immedi- 
ately placed in Miiller’s fluid for one week, then sectioned with 
the freezing microtome. The sections were mounted in glycerin 
for the study of the chromaffin reaction. The other gland was 
first fixed one or two hours in 10 per cent aqueous formalin solu- 
tion (neutralized with lithium carbonate). In a few cases the 
glands were left in 5 per cent formalin from eighteen to twenty- 
four hours, without apparent effect upon the liposomes. Bell 
(10), however, found in some cases a partial disappearance of 
the liposomes in muscle and epithelium after even a brief exposure 
to formalin. The formalin-hardened gland was cut with the 
freezing microtome into sections (at 10 y). 
Some of these sections were placed for about twenty hours in 
1 per cent aqueous osmic acid solution, and the others stained 
with scarlet red and mounted in glycerin for special study of the 
lipoids and fat. Herxheimer’s formula (saturated solution of 
scarlet red in 70 per cent alcohol containing 2 per cent of sodium 
hydroxide) was used. The sections were first placed in 60 per 
cent alcohol (one minute), then in Herxheimer’s fluid (three min- 
utes), then rinsed in 60 per cent alcohol (one-half minute), 
washed in distilled water a few minutes, and mounted in glycerin. 
Frozen sections were also studied fresh, mounted in normal 
salt solution, and also treated with 1 per cent potassium hydroxide, 
1 per cent acetic acid, alcohol, ete. The frozen sections were cut 
by means of a Spencer automatic freezing microtome, using 
liquid carbon dioxide for refrigeration. 
