466 METHODS AND FORMULAS DS 23.13 



23.13 Lenz 1907 23684, 44:374 



REAGENTS BEQUiRED: A. 0.5% eosin Y in 60% ale; B. water 100, potassium hydroxide 



0.1, sat. ale. sol. {circ. 2%) methylene blue 30; C. 0.005%, potassium hydroxide in abs. 



ale; D. 0.1% acetic acid in abs. ale. 

 method: [paraffin sections of acetone fixed and dehydrated material] -^ 70% ale. -> A, 



1 min. -* water, wash —> B, I min. -» blot or drain -* C, till yellowish -^ D, till nerve 



cells clear blue — > balsam, via usual reagents 



23.13 Lepine 1935 6630, 119:804 



REAGENTS REQUIRED: A. 0.5% magenta in 50% ale. 50, 0.2% safranin 50; B. abs. ale. 50, 



acetone 50; C. DS 11.44 Stevenel 1918 

 method: [paraffin sections of F 3700.0010 fixed material]-^ A, 10 mins. -^ B, quick 

 rinse -^ water, wash -^ C, 1 min. —>■ B, till section turns blue-^ water, wash ^ B, 

 quick dip -^ abs. ale, till section turns lilac -^ balsam, via xylene 



23.13 Lillie 1948 Lillie 1948, 225 



reagents required: A. abs. ale. 90, water 6, acetic acid 3.25, ethyl eosin 1; B. any 



DS 11.122 stain; C. 0.25% acetic acid 

 method: [alcohol-fixed material] -* .4, 2 mins. -^ 95% ale, rinse -^ water, wash ^ B, 

 till sufficiently stained -> C, till differentiated -^ balsam, via usual reagents 



23.13 McWhorter 1941 20540b, 16:143 



reagents required: A. any wetting agent; B. 0.5% phloxine; C. 0.9% sodium chloride; 



D. 0.5% trypan blue 

 method: [surface shaving from virus infected leaf] -^ A, brief rinse -^ B, 3-8 sees. -» C, 



brisk wash -^ D, 2-4 mins. — > C, for observation 

 result: viroplasts blue, red, or purple. 

 note: Formaldehyde-fixed material may be used after 1-6 hours in 10%, citric acid. 



Rich 1948 (20540b, 23:19) recommends pretreatment in water 85, sodium chloride 



0.7, 95% ale 10, ether 5. 



23.13 ManoueUan 1912 857, 26 :972 



reagents required: A. acetone 100, ADS 12.2Lugol (1905) 0.3; B. DS 13.7 Mann 1892 



(sol. yl); C. 2% glyceric ether in 95% ale 

 method: [fresh smears] -^ A, 5 mins. -^ acetone, thorough wash -^ B, 1 min. -^ C, till 



differentiated -* balsam, via usual reagents 

 note: Langeron 1949, 653 states that ADS 22.1 Beauverie and HoUande 1916, diluted 



50 : 1 with water may be used in place of glyceric ether. 



23.13 Nagle and Pfau 1937 617, 27 :356 



reagents required: A. phosphate buffer pH 7.4 90, 8% ale magenta 0.3, sat. sol. 



methylene blue 0.2 

 method : [dry impression smear] — » methanol, 2 mins. —> wash -^ B, on slide, heated to 

 steaming, 5 mins. -^ wash -^ dry 



23.13 Neri 1909 23684,50:409 



REAGENTS REQUIRED: A. watcr 100, iodinc 0.1, potassium iodide 0.2, eosin Y 1; 5. 1% 



methylene blue 

 PREPARATION OF a: Dissolve the iodine and iodide in a few drops of water. Dilute to 50. 



Add dye dissolved in 50 water to iodine solution. 

 method: [sections of acetone-fixed, or smears of alcohol-fixed material] -> A, 10-15 mins. 



-* water, wash -^ B, 5 mins. -* water, rinse -» 95% ale, till differentiated -> balsam, 



via usual reagents 



23.13 Petragnani 1928a test. 1930 Ciferri 20540b, 5:34 



REAGENTS REQUIRED: A. ADS 12.2 Petragnani (1928) (working sol.); B. 0.5% eosm B 



in 50% ale; C. DS 11.122 Mayer 1896; D. 0.1% methylene blue; E. 0.05% sodium 



hydroxide in abs. ale 

 method: [sections] -^ abs. ale -^ A, 5-10 sees. -^ abs. ale, rinse -^ B, 10-20 sees. -^ 



wash -^ C, 1 min. -^ rinse -^ D, till violet (not blue) -> blot -^ E, 15-20 sees. -^ 95% 



ale, till blue — > balsam, via usual reagents 

 result: Negri bodies, eosin-red; capillaries, nuclei, red; nerve cells, blue. 



