DS 23.12 DYE STAINS OF SPECIAL APPLICATION 463 



23.12 Hertig and Wolbach 1924 11343, 44:332 



REAGENTS REQUIRED: .4. DS 11.43 Goodpasture and Burnett 1919; B. 40% formalde- 

 hyde; C. sat. sol. picric acid 

 method: [thin sections of F 3700.0000 fixed material] —* A, poured on slide, warmed to 

 steaming, 5 mins. —>■ water, quick rinse -^ B, dropped on slide till no more color comes 

 away — > C, 1 min. -^ balsam, via usual reagents 



23.12 Laigret and Auburtin 1938 5310, 31:790 



PREPARATION OF DRT "salt": To a sat. sol. thionin add 10% sodium hydroxide until 



no further ppt. is formed. Filter. Wash and dry ppt. 

 WORKING solution: water 100, phenol 2, ppt. from above to sat. 

 method: [alcohol-fixed smears] — > stain, on slide, 30-50 sees. —* drain — > abs. ale, quick 



wash —> balsam, via xylene 



23.12 Lepine 1932 6630, 109:1162 



REAGENTS REQUIRED: A. Water 90, phenol 3, DS 11.43 Ziehl 1890 10; B. sat. aq. sol. 



{circ. 2.5%) Bismarck brown 

 method: [smears of vaginal scrapings, fixed on slide by F 7000.1000 Rdgaud 1910] — » 



water, wash -^ B, 2-3 mins. — ♦ water, wash -^ dry 

 result: Rickettsiae red in pale brown cells. 



23.12 Lepine 1933 6630, 112:17 



stock solutions: I. water 100, phenol 0.5, azur II 1; II. 1% potassium carbonate 

 REAGENTS REQUIRED: A. Water 100, stock I 3.5, stock II 2, 40% neutralized formalde- 

 hyde 5; B. 0.1% safranin 

 method: [very thin smears] — + ^, 3 or 4 mins. -^ water, wash — + B, till differentiated —^ 

 water, wash — > dry 



23.12 Macchiavello 1938 Macchiavello 1938, 48 



REAGENTS REQUIRED: A. sat. alc. sol. (circ. 4%) magenta 0.4, phosphate buffer pH 7.5 



100; B. water 100, thionin 0.01, citric acid 0.13; C. 0.5% citric acid 

 method: [thin smears]—^ A, on slide, 3 mins. -^ B, 2-3 sees. — > C, 2-3 sees. ^ tap 



water, thorough wash -^ dry 

 note: Zinsser, Fitzpatrick, and Hsi 1938 (11189, 69:179) omit step B and stain for 1 

 minute in 1% methylene blue after washing off the citric acid. This modification is 

 commonly referred to as " Macchiavello's method." The original method was cited by 

 Cox 1939 (17302, 63 :2242) to whom it is sometimes referenced. A detailed description 

 of the use of this technique is given under DS 23.10 above. 



23.12 Nyka 1944 11431,56:264 



reagents required: A. 5% magenta in 90% ale; B. 0.01% methyl violet; C. 0.007% 



acetic acid. 

 method: [3 M sections of F 7000.0000 Miiller 1859 fixed material] -^ 95 %o ale. -^ A, 5-10 



mins. — > rinse -^95% ale, 1-2 mins. -^ B, 1-3 mins. -^ rinse — ♦ C, till differentiated 



-> balsam, via acetone and xylene 



23.12 Nyka 1945 11431,52:317 



reagents required: A. 0.01% methyl violet; B. 0.05% acetic acid; C. 0.01% metanil 



yellow 

 method: [sections of formaldehyde-fixed material] —> water— > A, '^ to 1 hr. -^ B, till 



cytoplasm clear -^ rinse C, few sees. —> balsam, via acetone and xylene 



23.12 Roskin 1946 Roskin 1946, 158 



reagents required: A. water 80, 95% alc. 20, phenol 1, aniline 1, magenta; B. DS 11.44 



Loffler 1890 

 method: [sections] —> water —» il, 10 mins. —> wash -^ 95% alc. till differentiated—* 



wash —y B, 15-60 sees, —y wash — > abs. alc., minimum possible time — > balsam, via 



xylene 

 recommended for: Rickettsiae and Negri bodies. 



23.12 Stutzer 1911 23454,69:25 



reagents required: A. DS 11.44 Loffler 1890; B.\% tannin 



method: [thin sections] —» water —+'' A, 5-15 mins. —» rinse —> B, till differentiated-^ 

 wash — » blot -* abs. ale., least possible time — + balsam, via xylene 



