CHAPTER XXX. 385 



Wet films (ibid., 1909, p. 1751) are treated as follows : Fix them 

 for twelve to twenty-four hours in a mixture of 2 parts saturated 

 aqueous solution of sublimate with 1 of absolute alcohol. Wash 

 and treat for five to ten minutes with a mixture of 2 parts of iodide 

 of potassium, 100 of water, and 3 of Lugol's solution. Wash, and 

 treat for ten minutes with 0-5 per cent, solution of sodium thio- 

 sulphate. Wash, and stain as above (changing the stain for fresh 

 after half an hour), for one to twelve hours. Then pass through 

 mixtures of acetone with first 5, then 30, then 50 parts per cent, of 

 xylol into pure xylol, and mount in cedar oil. This process is 

 applicable to sections. 



Or (ibid., 1910, p. 2476) a slide is placed in a Petri dish and 

 covered with a mixture of equal parts of methyl-alcohol and stock 

 mixture. After half a minute this is poured off and enough distilled 

 water poured in to cover the slide, and the whole is rocked to mix 

 the two. After three to five minutes, wash in running water, dry, 

 and mount in cedar oil. 



By any of these processes nuclei (red) are demonstrated not only 

 in hsematozoa, but in many bacteria, spirochsetse, coccidia, sar- 

 cosporidia, etc. 



See also, for paraffin sections, SCHUBERG, in Deutsch. med. 

 Wochenschr., xxv, 1909, No. 48, or Zeit. wiss. Mik., xxvii, 1910, 

 p. 161, who passes through acetone and xylol into balsam. 



The older Romanowsky stains published by the authors men- 

 tioned above, as also Laveran's " Bleu Borrel " seem to be super- 

 seded by Giemsa's. 



IRISHMAN'S Romanowsky Stain (Brit. Med. Journ., March 16th 

 and September 21st, 1901) is as follows : To a 1 per cent, solution 

 of Grubler's medicinal methylen blue in water add 0-5 per cent, of 

 sodium carbonate, heat to 65 C. for twelve hours and let stand for 

 ten days. Then add an equal volume of 0-1 per cent, solution of 

 Griibler's Eosin extra B, let stand for six to twelve hours, collect 

 the resulting precipitate on a filter, wash it until the wash water 

 comes off colourless, dry and powder. For staining, dissolve 0- 15 grm. 

 in 100 c.c. of pure methyl alcohol. Stain cover-glass films (air- 

 dried) for five to ten minutes ; flood the film with water for one 

 minute, and examine, or dry (without heat) and mount in xylol 

 balsam. Nuclei in shades of red, cytoplasm bluish, parasites blue 

 with ruby red chromatin. 



RAADT (Munch, med. Wochenschr., 1911, No. 27 ; Zeit. wiss. Mik., 

 1912, p. 236) obtains a Romanowsky stain of blood and parasites 

 with JENNER'S solution. Films fixed with alcohol and ether are 



25 



M. 



