ZOOLOGY AND BOTANY, MICROSCOPY, ETC. 375 



this a considerable alcoholic firmness is obtained, and also a differentia- 

 tion of the nuclear and protoplasmic structure, and a staining of the 

 elastic fibres. Glanders bacilli [appear dark on an unstained ground, 

 and typhoid bacilli intensely red-violet. The proceeding is as follows : 

 (1) Fixing and hardening, best in Miiller-formalin, and embedding in 

 paraffin or celloidin. (2) Sections are stained overnight in orcein D 

 (Griibler), ' 1 ; officinal nitric acid, 2*0; 70 p.c. alcohol, 100 ' 0. (3) A 

 short washing in 70 p.c. alcohol. (4) Water. (5) Staining in poly- 

 chrome methylen-blue for 10 minutes to 2 hours. (6) Distilled water. 

 (7) Differentiation in glycerin-ether mixture, 1 ; water, 2 to 5, until the 

 sections appear bright blue. (8) Distilled water, 70 p.c. alcohol, abso- 

 lute alcohol, xylol, balsam. 



Is there a " Vital " Staining ? — Under this heading R. Krause* dis- 

 cusses the question as to whether vital staining is really possible, meaning 

 thereby a staining of the cell organs while the cells themselves suffer 

 no loss of function, or whether such staining is not merely staining 

 intra vitam, and associated with loss of function and approaching death 

 of the stained tissue elements. The author inclines to the former sup- 

 position from the results of observations on the ciliated cells lining the 

 vestibule of the labyrinth in Petromyzon, their function being of course 

 observable microscopically. He injected into the heart or posterior 

 cardinal vein of the living animal a few cubic centimetres of a 2 p.c. 

 solution of crystallised, chemically pure methylen-blue (Hochst) in 

 normal saline solution. At the end of the injection the auditory 

 capsule was laid bare, and then by means of a good knife horizontal or 

 vertical sections were cut. These were studied in normal saline solution. . 

 The cells at first appeared unaffected, but soon their constituent parts 

 underwent a differential staining, the continued unchanged movement 

 of the cilia contra-indicating any impairment in the functional activity 

 of the cells. 



Staining Trypanosoma. f — W. E. Musgrave and M. T. Clegg ap- 

 prove of "Woolley's method of staining Trypanosoma. The blood films 

 are fixed for 10 minutes in absolute alcohol, and then stained with the 

 following solution : — (A) Eosin, 1 grm. ; distilled water, 1000 c.cm. 

 (B) Polychrome methylen-blue, Unna's formula. (C) Methylen-blue, 

 1 grm. ; distilled water, 100 c.cm. (D) Solution B, 2 parts ; solution C, 

 1 part. 1 c.cm. of A is mixed with 4*5 c.cm. of D. The preparations 

 are stained for 20 to 40 minutes, are then washed, and afterwards stained 

 with solution A for 2 to 5 seconds. 



Method for Intra-vitam Staining of the Protoplasmic Granules 

 of the Cornea 4 — G. Colombo makes a saturated solution of Bismarck 

 brown in 92 p.c. sodium chloride. This is filtered while hot and after- 

 wards when cold. The solution is sterilised in a water bath and then 

 dropped into the conjunctival sac of a frog. About 5 drops are instilled 

 four times a day. In 3 or 4 days the cornea becomes yellowish brown. 

 A piece of the excised membrane may now be examined in physiological 

 salt solution. 



* Anat. Anzeig., xxiv. (1904) pp. 400-3. 



t Publications of Dep. Int. Bur. Govt. Lab., Manila, 1903. 



t Zeitschr. wiss. Mikrosk., xx. (1904) pp. 282-8 (1 pi.). 



