364 METHODS AND FORMULAS DS 13.51 



method: [sections of osmic-fixed, or mordanted, material] — > wash -^ A, overnight — > B, 

 till outline of nuclei distinct — > C, 20 niins. -^ B, till outline of nuclei distinct — » D, 

 till sections deep black, 1-3 mins. — > running water, till iodine removed — > E, till 

 sections bright blue, 1-3 mins. — > water, thorough wash —> blot — > 95%, rinse 5 sees. 

 — * carbol-xylene till differentiated, J£ to 5 mins. —>■ xylene, 2 mins. — > F, 1-2 mins. — > 

 clove oil wash -^ balsam, via xylene 



13.51 Henneguy 1898 test. 1907 Bohm and Oppel safranin-methyl violet-orange G 



Bohni and Oppel 1907, 111 

 REAGENTS REQUIRED: A. Water 100, ammonium thiocyanate 1, methyl violet 0.1, orange 



G 0.1; 5. DS 11.42 Zwaademaker 1887 

 method: [sections] -^ A, 10 mins. — > rinse -^ B, 15 mins. -^ rinse -^ A, 15 mins. — > abs. 



ale, minimum possible time — ^ balsam, via clove oil 

 result: nuclei, red; cytoplasm, various shades of blue and blue gray. 



13.51 Johansen 1940 safranin-methyl violet-fast green-orange G 



Johansen 1940, 88 

 REAGENTS REQUIRED: A. DS 11.42 Johanscu 1940; B. 1% methyl violet; C. 95% ale. 30, 



ethylene glycol monomethyl ether 30, tert. butyl ale. 30; D. clove oil 6, ethylene glycol 



monomethyl ether 6, fast green FCF q.s. to sat., 95% ale. 36, tert. butyl ale. 36, acetic 



acid 12; E. 95% ale. 50 tert. butyl ale. 50, acetic acid 0.5; F. sat. sol. orange G in 



ethylene glycol monomethyl ether 30, ethylene glycol monomethyl ether 30, 95% ale. 



30; G. clove oil 30, ethylene glycol monomethyl ether 30, 95% ale. 30; H. clove oil 30, 



abs. ale. 30, xylene 30 

 method: [sections] -^ 70% ale. -^ .4, 1-2 days -^ water, rinse ^^ B, 10-15 mins. — > 



water, rinse —> C, 15 sees. — * D, 10-15 mins. — * E, quick rinse -^ F, 3 mins. — > G, 



rinse — > H, rinse — > balsam, via xylene 

 result: (plant tissues) dividing chromatin, red; resting nuclei, purple: lignified and 



suberized tissues, red; cellulose, green-orange; cytoplasm, bright orange; starch 



grains, purple; fungal mycelia, green. 



13.51 Kalter 1943 safranin-crystal violet-fast green-orange II 



11284,28:995 

 REAGENTS REQUIRED: A. Water 50, 95% ale. 50, 40% formaldehyde 4, sodium acetate 

 0.5, safranin 0.2; B. 0.5% crystal violet; C. clove oil 100, fast green FCF and orange 

 II, each to sat.; D. sat. sol. orange II in clove oil 

 method: [sections] — > water — > A, 24 hrs. -^ rinse -^ B, 1-2 mins. -^ wash — » 95% ale. 

 -^ C, 5 mins. -^ clove oil, till connective tissue green -^ D, 10 mins. —* clove oil -^ 

 xylene, thorough wash — * balsam 

 result: nuclei, red; cytoplasm, pink or light green; muscle, tan; collagen, bright green. 



13.51 Laguesse 1901 test. 1907 Bohm and Oppel safranin-crystal violet-orange G 



Bohm and Oppel 1907, 154 

 REAGENTS REQUIRED: A. 2% potassium sulfite; B. DS 11.42 Babes 1887; C. sat. aq. sol. 



crystal violet 50, sat. aq. sol. orange G 2 

 PREPARATION OF C: Mix the solution. Add just enough water to redissolve ppt. 

 method: [sections] —> A, 12-24 hrs. -^ wash — > B, 6-12 hrs. ^ wash -^ C, 24 hrs. — > 

 blot — ^ abs. ale. minimum possible time — > clove oil, till differentiated —> balsam, via 

 xylene 



13.51 Stockwell 1934 safranin-grntial violet-orange G 19938,80:121 



REAGENTS REQUIRED: Aa. ADS 12.1 Stockwcll 1934 or Ab. 1% chromic acid; B.l% crys- 

 tal violet 20, 1% safranin O 40, water 40; C. AFS 12.1 LaCour 1931; D. 1% picric acid 

 in 95% ale; E. 95% ale. 100, ammon. hydroxide 0.3; F. 0.2%, orange G in clove oil 

 method: [sections]-^ water —> Aa, if bleaching required (or Ab if not chromic fi.xed) 

 overnight -^ water, thorough wash — » B, 1-6 hrs. — > water, rinse -^ C, 30 sees. -^ 

 70% ale, quick rinse -^ D, few sees. — » E, few sees. — > abs. ale, few sees. -^ F, few 

 sees. — ^ balsam, via xylene 



13.51 Unna 1928 test. 1928 Schmorl safranin-orcein-anilin blue-eosin 



Schmorl 1928, 343 

 stock solutions: I. water 50, abs. ale 25, glycerol 10, acetic acid 2.5, orcein 0.5, anilin 

 blue 0.5; II. 80% ale 100, ethyl eosin 1; III. 1%, hydroquinone 



