542 NEUROGLIA AND SENSE ORGANS 



100 c.c. and sublimate 3 grm., with the addition at the moment of use 

 of acetic acid 3 c.c, formalin 0-5 c.c. The fluid should be warmed at 

 35° to 40° C. and injected through the blood-vessels, the blood being 

 first washed away by means of Ringer's solution to which 1 : 1000 of 

 amylnitrite was added. The tissues are treated in the usual way and 

 imbedded in celloidin. The sections are first treated for five minutes 

 with a 1 per cent, solution of caustic soda in 80 per cent, alcohol and 

 washed in distilled water, and then mordanted for a few minutes in 5 

 per cent, iron alum and washed once more. For staining, Held adds 

 to some distilled water a few drops of a very old molybdic acid ha;ma- 

 toxylin, enough to impart to the water a bluish- violet tone, and stains 

 therein for twelve to twenty-four hours at 50° C. The stain is prepared 

 by dissolving 1 grm. of hsematoxylin in 100 c.c. of 70 per cent, alcohol 

 and adding an excess of molybdic acid. Differentiation is carried out by 

 means of the same iron-alum solution used for mordanting ; wash well ; 

 counterstain with v. Gieson picro-fuchsin solution ; wash in 96 per 

 cent, alcohol, dehydrate and mount as usual. 



Neuroglia cells and fibres greyish-black ; marginal neuroglia (mem- 

 brana limitans marginalis and membrana limitans perivascularis) sharply 

 differentiated ; connective tissue pink-red. Mallory's connective tissue 

 stain (acid fuchsin-anilin-blue-Orange G), and to a less extent Masson's 

 trichromic and M. Heidenhain's " Azan " stain (see § 811), give 

 useful neuroglia pictures. 



1089. Jakob (Nissl-Alzheimefs Arb. u. d. Grosshirnriude, v, 1913) 

 makes thin frozen sections of material fixed in gliabeize -t- 10 per cent, 

 formol, washes rapidly in distilled water, and stains first for three to ten 

 minutes in 1 in 1000 watery solution of acid fuchsin, and then after 

 washing puts into saturated watery solution of phosphomolybdic acid 

 for one to twenty-four hours. The sections are then, after another short 

 wash, put into a mixture containing 0-5 grm. of water-soluble anilin 

 blue, 2 grm. of Orange G, 2 grm. of oxalic acid and 100 c.c. of distilled 

 water, for half an hour After a short wash in distilled water the 

 sections are differentiated in 96 per cent alcohol, keeping them moving 

 in this, until no more clouds of stain come off. They are then treated 

 with absolute alcohol and xylol and motmted in balsam, (Glial cells 

 and processes violet, axis-cylinders blue, myelin sheaths golden with 

 slight rose tint. Ganglion cells and vessel walls dark blue, nuclei and 

 blood corpuscles light red.) 



Anderson (" Hozv to Stain the Nervous System,'''' E. and S. Livingstone, 

 Edinburgh, 1929), has modified this method for paraffin and celloidin 

 sections. Tissue to be imbedded in paraffin is best fixed in Bouin's or 

 Zenker's fluid, but if already fixed in formalin should be placed in 

 Bouin's fluid for sixteen to twenty-four hours before imbedding. This 

 treatment is not necessary for material to be imbedded in celloidin. 

 After washing in distilled water sections are placed for half an hour in 

 the following mordant: iron alum, 1 grm.; sulphuric acid, 1 c.c; 50 

 per cent, alcohol, 98 c.c The sections are then washed in distilled 

 water and stained for one to five minutes in |- per cent, acid fuchsin in 

 ^ per cent, glacial acetic acid. After a prolonged wash in distilled water 

 they are transferred to 4 per cent, phosphomolybdic acid for thirty to 

 forty-five minutes, washed again in distilled water and placed in 

 Mallory's anilin-blue-Orange G. mixture for fifteen minutes. Paraffin 

 sections are differentiated m absolute alcohol, celloidin sections in 95 

 per cent, alcohol. They are cleared in xylol and mounted in acid 

 balsam. 



Bailey (Journ. Med. Res., xliv, 1923, p. 73) cuts thin paraffin sections 

 of Zenker or Boain fixed material and after treatment with iodised 

 alcohol mordants for three days in 3 per cent. pot. bichromate, then 



