ANILIN CRYSTAL VIOLET 



25 



ANTHRAQUINONE DYES 



Filter and to 84 cc. of filtrate add 16 cc. 

 sat. ale. crystal violet. Leave 24 hrs. 

 before using. After about 10 days stain- 

 ing potency decreases (Mallory, p. 89). 



2. Stirling's. Crystal violet, 5 gm.; 

 abs. ale, 10 cc; anilin oil, 2 cc, aq. 

 dest., 88 cc. Keeps well (Mallory, p. 

 90). 



See Anilin Crystal Violet and Gentian 

 Violet. 

 Anilin-Fuchsin Methyl Green method for 

 mitochondria. This technique is based 

 on Altmann's method. It was used by 

 Bensley to stain tissues fixed in his 

 Acetic-Osmic-Bichromate fluid. Cow- 

 dry recommends instead fixation in the 

 better penetrating Regaud's fluid. 



Fix small pieces in freshly prepared 

 Regaud's fluid (3% aq. potassium bi- 

 chromate 4 parts, commercial formalin 

 1 part). Ordinarily it is not necessary 

 to neutralize the formalin before hand 

 by saturating it with magnesium car- 

 bonate. Keep in ice box and change 

 the fluid every day for 4 days. Pour 

 off fixative and mordant in 3% aq. po- 

 tassium bichromate 8 days changing 

 every second day. Wash in running 

 water over night or in several changes of 

 water. Dehydrate in alcohol, clear in 

 xylol, imbed in paraffin and cut sections 

 about 4 M thick. Pass mounted sec- 

 tions through xylol and alcohol to 

 water. Dry the slide with a cloth ex- 

 cept area covered by sections. Pour 

 on anilin acid fuchsin and heat to 

 steaming over a spirit lamp. (To make 

 this saturate 125 cc. aq. dest. with 

 anilin oil by shaking the two together. 

 Filter and add 15 gms. acid fuchsin 

 to 100 cc. of filtrate. Allow to stand 

 24 hrs. before using. It lasts about a 

 month.) Allow to cool and stain about 

 6 min. Pour stain back into bottle. 

 Remove most of remainder, except from 

 sections, with a cloth or filter paper. 

 Rinse in aq. dest. about 1 min. Allow 

 1% aq. methyl green, added with 

 a dropper, to flow over sections and 

 counter stain them. This usually takes 

 about 5 sec. but the time must be 

 determined by trial. Wash off excess 

 methyl green in 95% alcohol, dehydrate 

 quickly in absolute, clear in toluol 

 (or xylol) and mount in balsam. The 

 mitochondria are stained crimson and 

 the nuclei green. For colored illustra- 

 tions see Cowdry, E. V., Contrib. to 

 Embryol., Carnegie Inst, of Washing- 

 ton 1917, No. 11, 27-43. If the methyl 

 green does not stain intensely enough 

 treat the sections, before coloration with 

 fuchsin, with l%aq. potassium perman- 

 ganate 30 sec followed by 5% oxalic acid 

 30 sec. and wash in water. More methyl 

 green can be retained by blotting the 



sections after staining in it with filter 

 paper and by then passing directly to 

 absolute alcohol. If the time of fixation 

 and mordanting is reduced much below 

 that specified the fuchsin itself may not 

 color with sufficient intensity. Such 

 preparations hold their colors for a year 

 or more unless they have been unduly 

 exposed to sunlight, or the balsam is acid. 



Anilin Fuchsin Picric Acid, see Altmann's 

 method for mitochondria. 



Anilin Fuchsin Toluidine Blue and Aurantia, 

 see Champy-Kull method for mito- 

 chondria. 



Anilin Gentian Violet usually credited to 

 Ehrlich. Rarely is its composition 

 given exactly the same by any two 

 people. The "emended formula" (Soc. 

 Am. Bact.) is A: 2.5 gm. crystal violet 

 (85 per dye content) + 95% ethyl alco- 

 hol, 12 cc. B : anilin oil 2 cc. + aq. 

 dest. 98 cc. (shake, leave few minutes, 

 filter). Mix A and B. (McClung, 

 p. 137.) 



Anilin Oil. A good product is easily obtain- 

 able. It is much used in the making of 

 stains (cf. anilin fuchsin) and to clear 

 tissues from 95% alcohol and even sec- 

 tions from 70%. Lee (p. 71) says that 

 it should not be employed after fixation 

 in osmic acid and that unless removed 

 by chloroform or xylol it will give the 

 tissues and mounting medium a brown 

 coloration. 



Anilin Red, see Basic Fuchsin. 



Anilin- Safranin (Babes). Aq. dest., 98 

 cc. ; anilin oil, 2 cc ; excess of safranin O. 

 heat in flask in hot water bath at 70- 

 80 °C. Cool, filter and use filtrate. 



Anterior Chamber of Eye. This is in many 

 respects the best site for observations 

 on transplanted tissues. See trans- 

 plantation of uterine mucosa (Markee, 

 J. E., Contrib. to Embryol., Carnegie 

 Inst, of Washington, 1940, 28, 219-308) 

 and of tumors (Saphir, O., Appel, M. 

 and Strauss, H., Cancer Res., 1941, 1, 

 545-547). Aqueous humor is not so 

 species specific as other tissue fluids 

 that have been investigated. Conse- 

 quently transplants from other species 

 will often develop. There is of course 

 the advantage of direct observation 

 through the transparent cornea. More 

 recently the technique of transplanting 

 animal and human tumors into this 

 favorable environment has been de- 

 veloped mainly by Greene and his asso- 

 ciates. For discussion of literature 

 and techniques see Greene, H. S. N. and 

 Murphy, E. D., Cancer Research, 1945, 

 269-282. 



Anthraquinone Dyes. Derivatives of an- 

 thracene through anthraquinone. Acid 

 alizarin blue GR and BB, alizarin, 

 alizarin red S, purpurin. 



