454 TRANSACTIONS OF THE CANADIAN INSTITUTE. [VoL. VI 
sublimate reagents. The former was allowed to act for forty-eight 
hours, the material was then placed in seventy per cent. alcohol, which 
was replaced by a fresh quantity daily for a week, after which it was 
transferred to alcohol of ninety-five per cent. strength. The corrosive 
sublimate solutions were allowed to act for an hour only, and the sub- 
sequent treatment with alcohol was the same as in the case of the picric 
acid material. Material hardened in alcohol was used only for the iron 
and phosphorus reactions. 
The staining reagents found to be serviceable were Ehrlich’s and 
Delafield’s hematoxylin solutions, Czokor’s alum cochineal, safranin, 
eosin, picrocarmine and methylene blue. Material hardened with 
alcohol, picric acid or corrosive sublimate gave very valuable preparations 
when stained with picrocarmine solution for twenty-four hours, then for 
an hour with a dilute solution of hematoxylin. The picrocarmine 
specially selects the granules which Borzi and Palla term “ cyanophicin,” 
while the hematoxylin thus employed stains the “central body” and its 
granules particularly and the peripheral protoplasmic zone less dis- 
tinctly. The “central body” in corrosive sublimate preparations is less 
clearly differentiated from the peripheral zone. 
There is one advantage in the use of picric acid which does away with 
the necessity of resorting to transsections of the trichomes, for the reagent 
seems to have some solvent or disintegrating effect on the substance of 
the membranous sheath and of the transverse septa, the trichomes 
breaking up, under the slightest pressure of the cover-glass, into the 
separate cells which very frequently then are seen by their flat or end 
faces. 
The method of obtaining the reactions for “masked” iron in the 
Cyanophycez I have fully described elsewhere.' The iron, liberated by 
sulphuric acid alcohol, as indicated, was converted into Prussian blue, 
the trichomes were then stained with a picrocarmine solution for twenty- 
four hours when the cyanophycin granules acquired a deep red colour 
which contrasts markedly with the Prussian-blue tint of the ironholding 
granules. (Fig. 14). Instead of acid alcohol strong solutions of hydrogen 
peroxide which contained traces of sulphuric acid were frequently used 
to liberate the masked iron. 
The demonstration and localization of organic phosphorus were effected 
in the manner which I have indicated elsewhere,’ but I may here briefly 
1 ‘ The Distribution of Assimilated Iron Compounds, other than Hazmoglobins and Hzmatins, in 
Animal and Vegetable Cells,” Quart. Jour. Micro. Sci., Vol. XXXVIII, p. 175, 1895. 
2 ‘*On the Detection and Localization of Phosphorus in Animal and Vegetable Tissues,” Proceedings 
Roy. Soc., Vol. LXIII, p. 467, 1898. 
| 
. 
. de 
. 
. ‘ . P , 
SO eee se, Oe ee ee ee ee 
