GOODPASTURE'S METHOD 



114 



GRAM'S STAINS 



anilin oil, 1 cc; phenol crystals, 1 gm. 

 Wash in water. Differentiate in forma- 

 lin (37% solution of formaldehyde) few 

 seconds until bright red color changes 

 to rose. Wash in water. Counterstain 

 in sat. aq. picric acid .3-5 min. until 

 sections become purplish yellow. Wash 

 again in water. Differentiate in 95% 

 ale. until red reappears and some of it 

 as well as of the yellow is washed out. 

 Wash in water. Stain in Stirling's 

 gentian violet (gentian violet, 5 gms.; 

 95% ale, 10 cc; aniline oil, 2 cc; aq. 

 dest., 88 cc.) 5 min. or more. Wash in 

 water. Gram's iodine solution (iodine, 

 1 gm.; potassium iodide, 2 gms.; aq. 

 dest., 300 cc.) 1 min. Blot dry. Clear 

 in equal parts aniline oil and xylol until 

 no color is removed. Clear in 2 changes 

 xylol and mount in balsam. Gram- 

 negative bacteria, red; gram-positive 

 ones, blue; tissue red and blue; fibrin 

 deep blue. See his Polychrome Methyl- 

 ene Blue and Carbol-Anilin Fuchsin 

 Methylene Blue. 



Gordiacea, see Parasites. 



Gordon's SilverMethod. For blood smears, 

 also shows parasites, Gordon, H., J. 

 Lab. & Clin. Med., 1936-37, 22, 294- 

 298. Dry smears of blood or bone 

 marrow in air and fix in 10% formalin. 

 Wash in water. 2.5% aq. iron alum 

 10 min. or more. 4 changes aq. dest. 

 Dip in 1% aq. gelatin -|- 1 or 2 drops 

 2% sodium carbonate and drain. Wash 

 quickly in aq. dest. Impregnate 5-15 

 min. in silver solution (Add strong 

 ammonia drop by drop to 5 cc of 10.2% 

 aq. silver nitrate until ppt. is dissolved. 

 Add 5 cc. 3.1% aq. sodium hydroxide 

 and redissolve ppt. with strong 

 ammonia. With aq. dest. dilute to 100 

 cc). Wash in aq. dest. at 60°C. Re- 

 duce in: 10% formalin 90 cc. + 2.5% 

 iron alum 10 cc. Wash in tap water, 

 dehydrate in alcohol, clear in xylol 

 and mount in balsam. 



Gossypimine, see Safranin O. 



Grafts. Intracoelomic of eye primordium, 

 Joy, E. A., Anat. Rec, 1939, 74, 461- 

 486. See Transplantation. 



Gram's Iodine Solution. Iodine, 1 gm.; 

 potassium iodide, 2 gm.; aq. dest., 

 300 cc. A stronger solution may be 

 desirable with only 100 cc. aq. dest. 



Gram-Pappenheim stain as modified for 

 smears and paraffin sections (Scudder, 

 S. A., Stain Tcchn., 1944, 19, 39-44). 



Gram's Stains for bacteria : 



1. In smears. Hucker modification 

 (McClung, p. 138). Stain 1 min. in 

 equal parts A and B : A = crystal violet 

 (85% dye content, 4 gm.; 95% ale. 20 

 cc.) B = ammonium oxalate, 0.8gm.; 

 and aq. dest. 80 cc. After washing in 

 water immerse in: iodine, 1 gm. potas- 



sium iodide, 2 gm., aq. dest., 300 cc. 



1 min. Then wash in water and dry 

 by blotting. Decolorize 30 sec. in 95% 

 ale. gently moving. Blot and counter- 

 stain in : 10 cc. sat. safranin in 95% 

 ale and aq. dest. 100 cc. Wash and dry. 

 Kopeloff-Beerman Modification (Mc- 

 Clung, p. 139). Stain 5 min. or more 

 in : 1% aq. gentian or crystal violet, 1.5 

 cc. mixed before use with 0.4 cc. 5% 

 aq. sodium bicarbonate. Rinse in iodine 

 solution made by dissolving 2 gm. 

 iodine in 10 cc. normal sol. sodium 

 hydroxide and adding 90 cc. aq. dest. 

 and stand 2 min. or more. Blot dry. 

 Add 100% acetone drop by drop with 

 specimen tilted till no more color is 

 removed, less than 10 sec. Dry in air. 

 0.1% aq. basic fuchsin, 10-30 sec. 

 Wash in water and dry. Weiss Modifica- 

 tion (Weiss, E., J. Lab. & Clin. Med., 

 1940-41, 26, 1518-1519). Make thin, 

 uniform smears and fix over flame. 

 Cover slide with 3% gentian violet in 

 20% ale, 3-5 min. Wash in warm 

 water. Cover 3-5 min. with iodine, 20 

 gm. ; potassium iodide, 40 gm., aq. dest. 

 300 cc. Wash with warm water. De- 

 colorize in acetone and wash imme- 

 diately in water. Counterstain quickly 

 in 2% basic fuchsin in 95% ale. Wash 

 in water, dry and examine. 



The use of colloidal iodine has been 

 suggested to improve the reaction be- 

 tween bacteria and stain (Lyons, D. C, 

 J. Lab. & Clin. Med., 1936-37, 22, 

 523-524). Methods for preparing col- 

 loidal iodine are described by Chandler 

 and Miller (W. L. and E. J., J. Phys. 

 Chem., 1927, 31, 1091-1096). 



2. In sections. Gram-Weigert method 

 (McClung, p. 152). Fix in Zenker's 

 fluid. Stain paraffin sections lightly in 

 alum hematoxylin and wash in running 

 water. 1% aq. eosin, 1-5 min., followed 

 by washing in water. Stain |-1 hr. in 

 anilin methyl violet made by mixing 1 

 part of A with 9 of B : A. abs. ale 33 cc ; 

 aniline oil, 9 cc. ; methyl violet in excess. 

 B. Saturated aq. methyl violet and wash 

 in water. Lugol's iodine 1-2 min. and 

 wash in water. Blot; dehydrate and 

 clear in equal parts aniline oil and 

 xylol several changes. Wash with xylol 

 and mount in balsam. Glynn's method. 

 (Glynn, J. H., Arch. Path., 1935, 20, 

 896-899). To make stain triturate 1 

 gm. crystal violet and 1 gm. phenol 

 crystals in mortar and add 10 cc 

 absolute alcohol. Before using dilute 

 10 times with aq. dest., allow to stand 



2 days and filter. Stain deparaffinized 

 sections of Zenker (less acetic), Bouin, 

 Helly or 10% formalin fixed material 

 for 2 min. Drain off but do not wash. 

 Add Gram 's iodine , 1 min . Differentiate 



