346 METHODS AND FORMULAS DS 13.12 



13.12 Leishman 1901 3579,2:757 



PREPARATION OF DRY EOSiNATE: To 100 0.5% methylene blue add 0.25 sodium carbon- 

 ate Digest 12 hours at 65°C. followed by 10 days at 15°C. Filter. Add 50 0.5% eosin 

 B to filtrate. Leave 12 hours. Filter. Wash and dry filtrate. 



WORKING solution: methanol 100, dry stain 0.15 



method: [air-dried smear] —> stain, on slide, 5-10 mins. — > water, 1 min. — > blot dry 



note: This method is stated by Leishman {loc. cit.) to be a modification of a stain pro- 

 posed by Rowmanowski 1891 {St. Petersburg Med. Wschr., 16 :297) to the original of 

 which the writer has never had access. 



13.12 Lillie and Pasternack 1936 4349, 15:65 



preparation of dry stock: Dissolve 0.25 silver nitrate in 12.5 water. Add 1-2 5% 



sodium hydroxide, wash ppt. five times by decantation. Add 50 1% methylene blue to 



wet ppt. Shake at intervals for 11 days. Filter and add 45 1% eosin. Leave overnight, 



filter, and dry ppt. 

 formula of stock solution: methanol (acetone-free) 75, glycerol 25, dry stock 0.6 

 working solution: citric acid-sodium phosphate, dibasic buffer (see note) 70, acetone 



6, methanol 6, stock solution 4 

 note: The buffer must be so adjusted that after the addition of the other ingredients the 



pH is 5.3. 



13.12 Mallory test. Langeron 1942 Langeron 1942, 614 



reagents required: A. water 100, eosin Y 5; B. water 85, DS 11.44 Sahli 1885 15; 



C. ADS 22.1 Wolbach 1911 

 method: [water] -^ A, 20 mins. -^ rinse — > fi, 5 mins. -^ B, fresh solution, 15 mins. — » 



B, fresh solution, 30 mins. -* wash -^ C, till differentiated -^ abs. ale. least possible 



time — > balsam, via cedar oil 

 result: nuclei, blue; other structures, polychrome. 



13.12 Manwell 1945 11284,30:1078 



reagents required: A. DS 11.44 Manwell 1945; B. water, adjusted to pH 6.5 with 



acetic acid; C. 0.2% eosin Y 

 method: [methanol fixed smear] -* dry -^ A, 30 sees. -^ B, wash -> A, 30 sees. -^ dry 



— + neutral mountant 



13.12 Marie and Raleigh 1924 11284, 10:250 



preparation of dry stock: Dissolve 1 methylene blue in 100 0.5 sodium bicarbonate 



and expose to U.V. arc 30 mins. in shallow dish. Cool and add 500 0.1% eosin Y. 



Filter, wash, and dry ppt. 

 working solution: methanol 100, dry stock 0.16 



13.12 Michelson 1942 11284,27:551 



preparation of dry stock: Dissolve 1 methylene blue in 100 N/100 sodium hydroxide. 



Heat 2\i hours, 55°C. shaking for 1 minute at half-hour intervals. Add 1 sodium 



bromide, continue heating 2^2 hours. Cool. Filter. Add 60 1% eosin Y and mix. Add 



further eosin in doses of 5 till solution is reddish. Leave 24 hours. Filter. Dry filtrate. 



WORKING solution: 0.017% dry stock in methanol 



13.12 Raadt 1912 23632, 29:236 



STOCK solution: water 100, methylene blue 1, lithium carbonate 0.5 

 reagents required: .4. stock 10, water 90; B. DS 13.11 Jenner 1899 25, water 75 

 method: [methanol fixed smears] -^ A, on slide, 5-10 mins. -^ rinse -» blot -^ B. 5-10. 

 mins. — > wash -^ dry 



13.12 Roques and Jude 1940 /(.s7. 1949 Langeron Langeron 1949, 621 



formula of STOCK SOLUTION: methanol 50, glycerol 50, eosin B 0.2, methylene blue 0.2, 



DS 11.44 Roques and Jude 1940 0.5 

 preparation of STOCK SOLUTION: Dissolve dyes in methanol, leave 24 hours, filter, and 



add glycerol. 

 working solution: water 95, stock 5 



13.12 Rowmanowsky 1891 see DS 13.12 Leishman 1901 (note) 



