444 METHODS AND FORMULAS DS 22.21-DS 22.3 



22.21 Martinotti 1910 23632, 27 :24 



REAGENTS REQUIRED: A. water 75, lithium carbonate 0.5, toluidine blue 1.0, glycerol 20 



95% alcohol 5; B. 1% acetic acid 

 preparation: Dissolve in order given and ripen 1 month. 

 method: [distilled water] — » A, overnight —» B, till required inclusions differentiated. 



22.21 Milovidov 1928 6630, 98 :555 



REAGENTS REQUIRED: A. DS 22.21 Altmann (1920); B. 0.5% aurantia in 70% ale; C. 



water 100, phosphomolybdic acid 1, sodium hydroxide 0.1; D. DS 11.44 Unna 1892 



25, water 75 

 method: [paraffin sections of F 3700.1000 Milovidov 1928 fixed material] — » abs. ale. — > 



[varnish in thin collodion] -^ water — ^ A, 80°C., 1-3 mins. -^ wash — > B, few sees. — * 



wash —* D, few mins. — > rinse — > D, few mins. — > wash -^ balsam, via usual reagents 

 result: mitochondria red; bacteria blue. 

 recommended for: differentiation of mitochondria and bacteria in root nodules of 



legumes. 

 note: This was reprinted in English by Dufrenoy 1929 (20540b, 4:13) to whom it is 



often attributed. 



22.21 Parat 1926 4285a, 3 :222 



REAGENTS REQUIRED: A. DS 11.113 Kultschitzky 1889; B. ADS 21.1 Weigert 1885 

 method: [sections of F 3700.1000 fixed, and Parat 1926a (below) mordanted material] 

 -^ A, 24 hrs. 37°C. — ♦ B, till mitochondria differentiated 



22.21 Parat 1926a 4285a, 3 :220 



Any method is commonly referred to as Parat in which small pieces of fixed material 

 are treated with a sat. sol. potassium dichromate for 24-48 hours at 40°C. before being 

 sectioned and stained. 



22.21 Volkonsky 1928 4295a, 5 :220 



REAGENTS REQUIRED: A. DS 22.21 Altmann 1890, A sol.; B. 0.5% aurantia in 70% ale; 



C. 1% phosphomolybdic acid; D. DS 11.44 Unna 1892; E. DS 12.16 Unna 1895 

 method: [sections of F 3700.1000 Helly 1903 fixed and Parat 1926a treated material] -> 

 water — * A, on slide, heated to steaming, 1 min. — > B, till mitochondria differentiated 

 — *■ C, 1 min. — * water, wash — * D, 5-10 mins. -^ water, rinse — > E, till nuclei differ- 

 entiated — » balsam, via usual reagents 



22.21 Volkonsky 1933 test. 1942 Langeron Langeron 1942, 1105 



REAGENTS REQUIRED: A. Altmann 1890, A sol.; B. water 55, N/1 sodium hydroxide 10, 



phosphomolybdic acid 1, 95% ale. 35, aurantia 0.25; C. DS 11.44 Volkonsky 1933; D. 



DS 12.16 Unna 1895 

 PREPARATION OF B : Dissolve the acid in 30 water. Add the hydroxide. Dissolve the dye in 



25 water 35 ale. Add this solution to the first. 

 method: [sections prepared as for Volkonsky 1928 above] -^^ water —» ^, on slide, 



heated to steaming, 1 min. — » water, rinse — > B, till mitochondria differentiated — > 



water, rinse— > C, 10-15 mins. -^ water, rinse —> D, till nuclei differentiated-^ abs. 



ale., minimum possible time -^ balsam, via xylene 



22.21 Zimmermann 1896 see DS 22.5 Zimmermann 1896 



22.3 NissL Granules 



Nissl bodies were originally described by Nissl 1894, who used an alkaline solution of 

 methylene blue for their display. His^formula has generally been followed save that 

 toluidine blue has been substituted by most of the recent workers. Deipolli and Pomerri 

 1938 have substituted magenta for the blue dye in a method which will show mito- 

 chondria and bacteria as well as Nissl bodies. Attention should also be drawn to the 

 methods of Einarson 1932 and 1935 which use two of the oxazine dyes. The best method 

 forjdemonstration is, however, that of Windle, Rhines, and Rankin 1943 who have 

 worked out the satisfactory pH at which staining will take place after a series of given 

 fixatives. 



