452 CONNECTIVE AND ELASTIC TISSUES 



minutes more. A precipitate is formed. After cooling the 

 liquid is filtered, and the precipitate which remains on the filter 

 is brought back into the capsule, and there boiled with 200 c.c. 

 of 94 per cent, alcohol. Allow to cool, filter, make up the filtrate 

 to 200 c.c. with alcohol, and add 4 c.c, of hydrochloric acid. 



WoLFRUM {Zeit. wiss. Mik., xxv, 1908, p. 219) adds 10 to 15 

 per cent, of acetone to the mixture. 



Stain sections (of material fixed in any way) for twenty minutes 

 to an hour, wash with alcohol, clear with xylol (not with an 

 essence). Elastic fibres dark blue on a light ground. 



Crystal Violet for Weigert's Elastic Tissue Stain (Sheridan, 

 Journ. Tech. Meth. and Bull. Inter. Assn. Med. Museums, 1929). 

 Crystal violet 1 grm., resorcin 2 grm., distilled water 100 c.c. 

 Dissolve by boiling for a few minutes, stirring constantly. Add 

 to boiling mixture 30 c.c. of a 30 per cent, aqueous solution of ferric 

 chloride. Continue boiling, stirring constantly, for several minutes 

 or until precipitation ceases. Collect precipitate on the filter and 

 wash with 50 c.c. of distilled water. Dissolve in 100 c.c. of 

 absolute alcohol by boiling in a porcelain capsule on a sand-bath. 

 Cool, filter and add 2 c.c. HCl. Stain sections for one or two 

 hours and differentiate in absolute alcohol till elastic fibres alone 

 remain. 



MiNERViNi (Zeit. wiss. Mik., xviii, 1901, p. 161) gives a variant with 

 safranin instead of fuchsin. 



See also Pranter, ibid., xix, 1903, p. 361 ; B. Fischer, Virchow's 

 Arch., clxx, 1902, p. 285, or Zeit. wiss. Mik., xx, 1903, p. 40 (chemistry 

 of the dyes obtained by these processes, which he calls " Fuchselin," 

 " Safranelin," etc.) ; Hart, Centralb. allg. Path., xix, 1908, p. 1 ; and 

 CiLiMBARis, Arch. mik. Anat., Ixxv, 1910, p. 708. 



974. Haematoxylin Methods. Harris {Zeit. wiss. Mik., xviii, 1902, 

 p. 290) makes an *' Elasthaematein " as follows : Haematoxylin 0-2 grm., 

 aluminium chloride 01 grm., alcohol of 50 per cent. 100 c.c, boil and 

 add mercuric oxide 0-6 grm., filter and add 1 drop of HCl. Keep for 

 some weeks. Stain for five or ten minutes, put into alcohol with 1 per 

 cent, of nitric acid for one minute, then pure alcohol. 



See also De Wit, Anat. Rec, i, 1897, p. 74 ; Duerck, Arch. Path. 

 Anat., clxxxix, 1907, p. 62 ; Verhoeff, Journ. Amer. Med. Assoc, 1908, 

 No. 11. 



Mallory's phosphotungstic haematoxylin is good, but not specific. 



For a haematoxylin and eosin stain for connective tissues see Kruger 

 (Zeit.f. IV. Mikr., xxxi, or Journ. R. Micr. Soc, 1914). 



975. Other Methods for Elastic Tissue. For the elastic tissue of the 

 skin see Passarge and Krosing, Derm. Stud., xviii, 1894. 



See also for staining and dissociation Agababow, Arch. mik. Anat , 1, 

 1897, pp. 566 et seq. 



For C. Martinotti's silver impregnation see Zeit. wiss. Mik., v, 1888, 

 p. 521, or Arch. Ital. Biol., xi, 1889, p. 257. 



Schumacher {Arch. mik. Anal., Iv, 1899, p. 151) has had good results 

 (for the spleen) with picro-nigrosin. 



976. Drew-Murray van Gieson-Nile Blue Method for Con- 

 nective Tissues (and Bacteria). Fix in formol-salt solution. 



