548 METHODS FOR INVERTEBRATES. 



bleaches if necessary with hydrogen peroxide, and stains with 

 picrocarmine or Delafield's hsematoxylin. He also embeds in 

 paraffin, 1022, and stains sections with iron hsematoxylin. 



ZACHARIAS (Zool. Anz., xxii, 1899, p. 72) fixes Uroglena, etc., with 

 a mixture of 2 volumes saturated aqueous solution of boracic acid 

 and 3 of saturated sublimate. 



1034. Stains for Flagella. The ROMANOWSKY stain will give a red 

 stain of the flagella of some forms. 



The method of LOFFLER (Centralbl. BakterioL, vi, 1889, p. 209 ; 

 vii, 1890, p. 625 ; Zeit. wiss. Mik., vi, 1889, p. 359 ; vii, 3, 1890, 

 p. 368 ; Journ. Roy. Mic. Soc., 1889, p. 711 ; 1890, p. 678) is as 

 follows. To 10 c.c. of a 20 per cent, solution of tannin are added 

 5 c.c. of cold saturated solution of ferrous sulphate and 1 c.c. of 

 (either aqueous or alcoholic) solution of fuchsin, methyl violet, or 

 " Wollschwarz." Cover-glass preparations are made and fixed in 

 a flame in the usual way, special care being taken not to over-heat. 

 Whilst still warm the preparation is treated with mordant (i.e. the 

 above- described mixture), and is heated for half a minute, until the 

 liquid begins to vaporise, after which it is washed in distilled water 

 and then in alcohol. It is then treated in a similar manner with the 

 stain, which consists of a saturated solution of fuchsin in anilin 

 water (p. 177), the solution being preferably neutralised to the point 

 of precipitation by cautious addition of 04 per cent, soda solution. 



See also LIEBETANZ, Arch. Protistenk., xix, 1910, p. 23. 



BUNGE (Journ. Roy. Mic. Soc., 1894, p. 640 ; Zeit. wiss. Mik., 

 xiii, 1896, p. 96) makes the mordant by mixing 3 parts of the tannin 

 solution with 1 of liquor ferri sesquichlorati diluted twentyf old with 

 water, and lets the mixture ripen for some days exposed to the air, 

 or (Journ., 1895, pp. 129, 248) adds to it a few drops of hydrogen 

 peroxide, until it becomes red-brown, when it is shaken up and 

 filtered on to the cover-glass and allowed to act for a minute. The 

 cover-glass is then mopped up and dried, and stained with carbol 

 gentian. 



KOERNER and FISCHER (quoted from Encycl. mik. Techn., p. 514) 

 make the mordant with 2 parts of tannin, 20 of water, 4 of ferrous 

 sulphate solution of 1 : 2 strength, and 1 of saturated alcoholic 

 solution of fuchsin. Warm, let it act for a minute, rinse and stain 

 with anilin-water-fuchsin, or carbol fuchsin. 



Similarly ELLIS (Centralb. Bakt., xxi, 1903, p. 241 ; Journ. Roy. 

 Mic. Soc., 1904, p. 249), but staining with Saureviolett, 1 part to 75 

 of alcohol and 75 of water. 



PEPPLER (Centralb. Bakt., xxix, 1901, p. 376 ; Zeit. wiss. Mik., 



