510 METHODS AND FORMULAS DS 23.33 



23.33 Shortt 1927 9940, 14:565 



fokmula: xylene 75, phenol 25, eosin Y 1 

 preparation: Dissolve dye in phenol. Dilute with xylene. 

 method: [fixed smears, either dehydrated in ale, or air dried] —> stain, 1-5 mins. -> 



xylene, thorough wash -^ balsam 

 RECOMMENDED FOR: demonstration of parasitic Sarcodina, and their cysts, in smears. 



23.33 Simons 1938 5310, 31:100 



REAGENTS REQUIRED: A. Water 100, methylene blue 0.2, sodium chloride 0.6, sodium 



citrate 1, saponin 0.6, 6% formaldehyde 4; B. 50% glycerol 

 PREPARATION OF A: Dissolvc the dye and sodium salts in water with gentle heat. Cool. 



Add saponin and shake till dissolved. Add formaldehyde. 

 method: [mix on slide 1 part blood to 5 parts A] — * dry — > B, tiU differentiated -^ seal 



coverslip 

 RECOMMENDED FOR: staining Plasmodium in fresh blood. 



23.33 Sinton and MulUgan 1930 9940, 17 :329 



REAGENTS REQUIRED: A. 0.04% sodium hydroxide in 90% ale; 5. 1% Bordeaux red; 



C. 4% ferric alum; D. DS 11.111 Shortt 1923 (sol. B); E. 0.3% iron alum; F. 0.1% 

 eosin 



method: [thick smears fixed in F 0000.0010 Gilson, 1897]^ 50% ale. -> A, several 

 hours -^ water, via graded ales. -^ B, 12-24 hrs. — ♦ rinse — > C, 18 hrs. -* rinse -^ D, 

 overnight — > E, till differentiated — * F, 3 mins. — > wash -^ balsam, via usual reagents 



recommended for: Plasmodium in thick smears. 



23.33 Sternberg 1905 23681, 16 :293 



reagents required: A. DS 13.13 Giemsa 1902 3, water 97; B. 0.5% 5% acetic acid 

 method: [thin paraffin sections] — > water ^ A, 24 hrs. -^ wash — > B, till section red — > 



wash — > dry -^ abs. ale, till blue — > dry — * balsam 

 recommended for: Plasmodium in sections. 



23.33 Tomlinson and Grocott 1944 591b, 14:36 



reagents required: A. water 100, calcium chloride 0.01, phloxine 1; B. water 75, 

 lithium carbonate 0.5, toluidine blue 1, glycerol 20, abs. ale. 5; C. 0.2% acetic acid; 



D. acetone 75, abs. ale. 25, rosin 3.75, orange G 0.01 



method: [sections] — > water -^ A, b mins. -^ rinse -^ B, 45-60 sees. — > rinse -^ acetone, 

 2 mins. -^ C, till blue clouds cease — » D, 1-2 mins. — » balsam, via acetone and xylene 

 recommended for: Plasmodium in sections. 

 note: Pigmented tissues may be bleached in 5% ammonium sulfite before this stain. 



23.33 Tompkins and Miller 1947 591b, 17:755 



reagents required: A. 4% ferric alum; B. 0.5% hematoxylin; C 2% phosphotungstic 



acid. 

 method: [smears fixed in F 3000.0000 Schaudinn 1893 after iodine treatment for re- 

 moval of mercury] -^ water -^ A, 3-5 mins. -^ rinse— > B, 1 min. — * rinse—* C, 2 

 mins. — > blue in tap water or alkali — * wash — > dammar, via usual reagents 

 recommended for: protozoans in fecal smears. 



23.33 Tsuchiuya 1932 11284, 17:1163 



reagents required: A. 4% ferric alum; B. DS 13.12 Wright 1910 

 method: [fecal smears fixed in F 3000.0000 Schaudinn 1893] — > A, 20 mins. — > wash, 3 

 mins. — > B, on slide, 1 min. — > B, diluted on slide, 5 mins. — > wash — * balsam, via 

 usual reagents 

 recommended for: intestinal protozoans. 



23.33 VUlain and Comte 1933 1843, 22 :137 



stock solutions: I. 0.08% eosin B; II. 0.08% azur II 

 working solution: stock I 6.5, stock II 6.5, water 87 

 method: [alcohol-fixed smears] — > stain, }4 to 1 hr. — > wash — » dry 

 note: All solutions must be adjected to pH 7.4 to 7.6. I 



recommended for: Plasmodium. 



