402 METHODS AND FORMULAS DS 21.211 



method: [A, by copious injections into the living animal] -^ [small pieces of brain] — > B, 

 15 mins. — > C, 4-12 hrs. -* wash -^ [paraffin, via usual reagents] — » sections -^ any 

 DS 11.21 formula if counterstain required 



note: Bethe, loc. cit., gives the following alternative formulas for C: I. water 100, am- 

 monium molybdate 5, hydrochloric acid 0.2; II. water 100, ammonium molybdate 5, 

 chromic acid 1, hydrochloric acid 0.2; III. water 100, ammonium molybdate 5, osmic 

 acid 0.25, hydrochloric acid 0.2; IV. water 100, sodium phosphomolybdate 5, osmic 

 acid 0.25, hydrochloric acid 0.2; V. water 100, sodium phosphomolybdate 5, hydro- 

 chloric acid 0.2. Cole 1933 (20450b, 9, 89) prefers: water 50, glycerol 50, hydrochloric 

 acid 0.5, ammonium molybdate to excess. A detailed description of the use of this 

 technique is given under DS 21.20 above. 



21.211 Bethe lest. 1933 Cajal and de Castro Cajal and de Castro 1933, 169 



REAGENTS REQUIRED: .4. 5% nitric acid; B. G5% ale. 90, ammonium, hydroxide 10; C. 

 70% ale. 90, hydrochloric acid 10; D. 4% ammonium molybdate; E. 0.03% toluidine 

 blue 

 method: [small fragments of fresh tissue] -^ A, 24 hrs. — > 95% ale, 24 hrs. — > 5, 1 wk. 

 -^95% ale, thorough wash -^ C, 24 hrs. ^95% ale, thorough wash -^ water, 

 wash -^ D, 24 hrs. -^ [5-8 m paraffin sections] -^ water, 10-12 mins., 60°C. — * E, 1-10 

 mins. — > water, till differentiated — > abs. ale, till dehydrated — * balsam, via xylene 

 RECOMMENDED FOR: neurofibriUae. 



21.211 Bethe and Monckeberg test. 1933 Cajal and de Castro 



Cajal and de Castro 1933, 335 

 reagents required: A. 0.25% osmic acid; B. water 100, sodium bisulfite 2, hydro- 

 chloric acid 1; C. 2.5% ammonium molybdate; D. 0.1% toluidine blue 

 method: [pieces of nerve] -> A, 24 hrs. -^ wash, 6 hrs. -^ 96%, ale, 24 hrs. -> wash, 4 

 hrs. — > B, 18 hrs. -^ wash, 2-4 hrs. -* [2 m-3 m paraffin sections] -^ water — » B, 5-10 

 mins. 20°-30°C. — > rinse -^ C, 5 mins. 50°-60°C. — > rinse -^ balsam, via usual 

 reagents 

 recommended for: neurofibriUae in medullated fibers. 



21.211 Bing and EUerman 1901 1739, 3 :260 



REAGENTS REQUIRED: A. sat. sol. (circ. 4.5%) methylene blue; B. sat. sol. (circ. 1.2%,) 



picric acid 

 method: [hand sections of Bing and Ellerman 1901, F 0000.1300 fixed material] -♦ A, 



5-10 mins. -^ B, 1 to 2 mins. -^ acetone, till dehydrated -^ balsam, via xylene 



21.211 Cajal 1893 test. Pollack 1900 Pollack 1900, 133 



REAGENTS REQUIRED: A. methylene blue (solid powder); B. 0.1% hydrochloric acid; C. 



water 100, ammonium molybdate 10, hydrochloric acid 0.3; D. F 2000.1000 Cajal 



1893; E. 0.3% platinic chloride; F. 0.3%, platinic chloride in abs. ale 

 method: [small pieces of fresh, or living, tissue] -^ A, dusted over surface, 45 mins. -^ 



B, quick wash ^ C, 2 to 3 hrs. -* rinse -^ D, 3-4 hrs. --' wash briefly -^ E, 15 mins. 



— > F, till dehydrated -^ [paraffin sections] -> balsam, via usual reagents 



21.211 Chang 1936 763, 65:437 



formula: water 90, 40% formaldehyde 10, thionin 0.3 



method: [fresh brain tissue] — > stain, few days to few months -^ wash -^ [paraffin sec- 

 tions] -^ balsam, via usual reagents 

 result: fiber tracts, red; cell bodies, blue. ■^ 



21.211 Cole 1933 see DS 21.211 Bethe 1896 (note) 



21.211 Donaggio test. 1933 Cajal and de Castro Cajal and de Castro 1933, 72 



REAGENTS REQUIRED: A. pyridine; B. water 100, ammonium molybdate 4, hydrochloric 



acid 0.2; C. 0.01% thionin 7 

 method: [small fragments] -^ A, 5-6 days -* wash, 24 hrs. -^ 5, 8 days -^ wash -^ 



[5 M paraffin sections] -^ C, till sufficiently stained -^ quick rinse -^ B, b mins. -^ 



balsam, via usual reagents 

 recommended for: neurofibriUae. 



