GOMORI'S METHODS 



148 



GORDON'S SILVER METHOD 



for U to 24 hours at 37°C.: 



Molar acetate buffer pH 5* 2.5 ml. 



5% lead nitrate 1.5 ml. 



Dist. water 40.0 ml. 



2% Na glycerophosphatet 7.5 ml. 



• 100 cc. of 13.6% CHiCOONa-3HiO plus 60 cc. 6% 



acetic acid, 

 t Commercial grade (mixture of alpha and beta salts) • 



Shake well, heat to ±60°C. for about 

 10 min.; filter. 



8. Rinse sections thoroughly first in 

 dist. water and afterwards in 2 to 3% 

 acetic acid, followed again by dist. 

 water. 



9. Immerse sections in a solution of 

 yellow ammonium sulfide (1-12 drops to 

 a Coplin jar) for 1 minute. 



10. Wash. Counterstain as desired. 

 For Alkaline phosphatase: 



1. Fix thin slices of tissues in 80% al- 

 cohol (or absolute acetone). Dehy- 

 drate in 95% and absolute alcohol (or 2 

 changes of absolute acetone), embed 

 through benzene or xylene in paraffin. 

 Cut sections around 6 micra thick. 



2. Run slide through xylene and 2 al- 

 cohols to distilled water. Incubate for 

 1 to 2 hr. at 37°C. in the following mix- 

 ture: 



2% sodium glycerophosphate. 25 cc. 

 2% sodium barbital 25 cc. 



Distilled water 50 cc. 



2% calcium chloride 5 cc. 



2% magnesium sulfate 2 cc. 



Chloroform a few drops 



This solution will keep in the ice box 

 for months. 



3. Rinse slide thoroughly in repeated 

 changes of distilled water. 



4. Immerse slide for 3 minutes in a 

 1 to 2% solution of some cobalt salt 

 (chloride, acetate, sulfate). 



5. Wash thoroughly under the tap. 



6. Immerse slide for 2 minutes in a 

 dilute solution of yellow ammonium sul- 

 fide (1-12 drops to a Coplin jarful of 

 distilled water). Wash under the tap. 



7. Counterstain as desired; dehy- 

 drate, clear and mount. 



Attention is called to the earlier 

 demonstration of phosphatase in bone 

 by Robison (R., Biochem. J., 1923, 17, 

 286-293) and to recent discussion by 

 Blaschko and Jacobson (Bourne, pp. 

 217-221). The distribution of phos- 

 phatase in some normal tissues is indi- 

 cated in colors by ICabat, E. A. and 

 Furth, J., Am.J. Path., 1941, 17, 303-318. 

 For phosphatase in elementary bodies 

 of vaccinia virus, see Macfarlane, 

 M. 0., and Salaman, M. H., Brit. J. 

 Exp. Path., 1938, 19, 184; Hoagland, 



C. L. et al., J. Exp. Med., 1942, 76. 

 163-173. See Kidney. 

 Gonococcus, methyl green-pyronin stain. 

 To 10 cc. absolute methyl alcohol add 

 1 gm. methyl green (dye content_60%) 

 and 0.2 gm. pyronin (bluish certified). 

 Add 100 cc. 2% aq. phenol and shake 2 

 hrs. per day for 2 days in a mechanical 

 shaker. Filter and add 20 cc. glycerin, 

 C.P. to filtrate. Fix smears by passing 

 slides lengthwise through flame 4 or 5 

 times. Add stain immediately and warm 

 to slight steaming. Wash off stain 20-50 

 sec. Dry and examine. Gonococci, 

 deep red ; other bacteria except these of 

 Neisseria group pale purplish or barely 

 noticeable; nuclei of pus cells green in 

 soft pink or rose cytoplasm (Walton, 

 S. T., J. Lab. & Clin. Med., 1938-39, 

 24, 1308-1309). 

 Goodpasture's Method as modified by Mac- 

 Callum for bacteria in sections 

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

 fluid or in formalin Zenker. Stain thin 

 paraffin sections 10-30 min. in: 30% 

 ale, 100 cc; basic fuchsin, 0.59 gr.; 

 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. 



Gopal-Ayengar, see Chromosomes, Hyal- 

 uronic Acid. 



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 -f 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% 



