CHAPTER XXXII. 415 



NISSL'S remarks thereon, Deutsche Ztschr. Nervenheilk, xiii, 1899, p. 348 ; 

 ILBERG, Neural Centrbl, xv, 1896, p. 831 ; Cox, Anat. Hefte, x, 1898, 

 p. 75 ; Int. Monatschr. Anal,x.v, 1898, p. 241 ; AUERBACH, Monatschr. 

 Psych. Neurol, iv, 1898, p. 31 ; MYERS, Anat. Rec., ii, 1908, p. 434 ; 

 SAVINI, E. u. TH., Centrbl Bakl, I Abth., xlviii, 1909, p. 697 ; MOSSE 

 (argentamin stain), Arch. mikr. Anal, lix, 1902, p. 403 ; MENTZ v. 

 KROGH, Centrbl Bakl, I Abth., Iviii, 1911, p. 95 ; JOHNSTON, Anal Bee., 

 xi, 1916, p. 287. 



832. HELD'S Methylene Blue and Erythrosin Method (Arch. Anat. 

 Phys., Anat, Abth., 1895, p. 399 ; 1897, pp. 226233, 273385, 

 Supplementband). Material may be fixed in alcohol, but preferably 

 either in picro-sulphuric acid, or in van Gehuchten's mixture of 

 alcohol, chloroform and acetic acid, or 1 per cent, corrosive sublimate 

 in 40 per cent, acetone. Tissues should be carefully embedded in 

 paraffin and sections stuck to slides by the water method. They are 

 stained with the aid of gentle heat for one to two minutes in a 

 solution of 1 grm. erythrosin in 150 of distilled water acidulated with 

 2 drops of glacial acetic acid. After washing with water the slides 

 are transferred into a mixture of equal parts of Nissl's methylene 

 blue solution and 5 per cent, acetone, warming until all odour of the 

 latter has disappeared. Differentiation is carried out after cooling 

 by means of a -1 per cent, solution of alum until sections are reddish. 

 Rinse in distilled water, dehydrate as rapidly as possible in absolute 

 alcohol, wash in xylol and mount in balsam. 



BOCCARDI (Monit. Zool. Ital, x, 1899, p. 141) uses a mixture of 

 erythrosin 0-1 grm., toluidine blue 0-2 grm., and water 100 c.c., and 

 differentiates in -5 per cent, alum solution. 



By means of Held's method, besides the tigroid substance, other 

 granules viz., Held's neurosomes become stained. It may, there- 

 fore, be considered as a typical example of double staining of nerve- 

 cells. Other double stains demonstrating basophil, acidophil and 

 other granules have been repeatedly proposed and may be easily 

 obtained by the combination of an acid and a basophil dye. One 

 generally uses watery solutions, e.g., of acid fuchsin, and methylene 

 or toluidine blue, and one stains first with the acid dye and then with 

 the basophil one, differentiation being carried out with alcohol. 

 One may also have recourse to EHRLICH'S triacid ( 296) as originally 

 proposed by ROSIN (Neurol. Centrbl., xii, 1893, p. 803), or to one or 

 other of the methods used for staining blood films ( 784), such as 

 PAPPENHEIM'S panoptic triacid stain, Jenner's mixture, Leishman's 

 Romanowsky stain, Pappenheim's method as described in 785, 

 and so on. 



See on this subject COWDRY, Int. Monatschr. Anat., xxix, 1913, 



