MS 33.21 METAL STAINS 581 



33.2 Neurological Methods 



33.21 nerve cells and processes 



33.21 Agduhr 1917 23632,34:1 



REAGENTS REQUIRED: A. 20% neutralized formaldehyde; B. 3% silver nitrate; C. MS 



33.1 Agduhr 1917; D. 0.5% acetic acid; E. 0.004% gold chloride; /''. 5% sodium 



thiosulfate 

 method: Fix .1, 5 days -^ wash -^ B, G days in dark — ♦ wash —> C, 20 hrs. -+ D, 1 hr. -> 



wash, 1 hr. -^ .4, 1 to 4 days —> [section by paraffin technique, bring sections to water] 



-^ E, 1 hr. -^ F, 2 mins. -^ balsam, via usual reagents 

 note: Agduhr emphasized the need for thorough washing, particularly between A and 



B and for the use of large volumes (100 ml. for 5 nun. cubes) of A. 

 recommended for: neurofibrils. 



33.21 Bielschowsky 1904 method for pieces — auct. 11478,3:109 



reagents required: A. 8% neutralized formaldehyde; B. pyridine; C. 3% silver 

 nitrate; D. MS 33.1 Bielschowsky 1902 40 ml., water 60 ml.; E. 0.004% gold chloride; 

 F. 5% sodium thiosulfate 

 method: [Fix A, 1 month or till required] -^ B, 3 days -* running water, 24 hrs. — > dis- 

 tilled water, wash -» C, 3 days, 36°C. -> rinse -^ D, 24 hrs. -^ wash -^ A, 10 hrs. -^ 

 [sections by paraffin technique, bring sections to water] —> E, 1 hr. -^ F, 2 mins. -* 

 balsam, via usual reagents 



33.21 Bielschowsky 1905 see MS 33.21 Bielschowsky 1910 



33.21 Bielschowsky 1908 test. 1933 Cajal and de Castro 



Cajal and de Castro 1933, 365 

 reagents required: A. 8% formaldehyde; B. 5% nitric acid; C. 4% silver nitrate; D. 



MS 33.1 Bielschowsky 1902; E. 0.2% gold chloride; F. 5% sodium thiosulfate 

 method: [fresh tissue] — > A, till hardened -+ B, till decalcified — > wash, till acid-free — > 

 A, 24 hrs. — > wash — > [frozen sections]-^ water ^^ C, 24 hrs. in dark -^ rinse -^ D, 

 till brown, 20 mins. -> wash -> A, 10-15 mins. -^ rinse -^ E, till grey -^ F,b mins. -♦ 

 balsam, via usual reagents 

 recommended for: nerve endings in structures requiring decalcification. 



33.21 Bielschowsky 1910 method for sections — auct. 11478,3:169 



REAGENTS REQUIRED: A. 2% silver nitrate; B. MS 33.1 Bielschowsky 1904; C. 8% 

 neutral formaldehyde; D. 0.004% gold chloride; E. 5% sodium thiosulfate 



method: [sections by freezing technique, of formaldehyde-fixed material, from which 

 the formaldehyde must be completely washed out before sectioning] -^ A, 24 hrs. in 

 dark —>■ rinse — > B, 15 to 20 mins. — > wash — » C, 15 to 30 mins. —> wash — > D, 1 hr. — » 

 E, 2 mins. — > balsam, via neutral reagents 



notes: Bielschowsky 1905 (11478, 4:227) varies from above in substituting a few 

 seconds in 2% acetic acid for the wash between B and C above. Favorsky 1906 (11478, 

 6:260) substitutes 10% silver nitrate for A above. Boeke 1910 (766, 35:193) substi- 

 tutes his F 0000. 1000 for plain formaldehyde in fixation ; otherwise his method is es- 

 sentially that above 



RECOMMENDED FOR: peripheral nerve endings. 



33.21 Boecke 1910 see MS 33.21 Bielschowsky 1910 (note) 



33.21 Cajal 1925 21344,23:157 



reagents required: A. AMS 11.1 Cajal 1913; B. MS 33.1 Cajal 1925 33.1; C. 4% neu- 

 tralized formaldehyde; D. 0.2% gold chloride; E. 5% sodium thiosulfate 



method: [sections by freezing technique of materials hardened 5-30 days in A] — > A, 4 

 hrs. -^ wash -^ B, warming if necessary, till dark amber -> wash — » C, 1 min. -^ D, 

 till required color —* E, 2 mins. — > balsam, via usual reagents 



note: This technique is frequently confused with MS 31.21 Cajal 1925. 



RECOMMENDED FOR: Cerebellum. 



