GOLGI METHODS 



113 



GOODPASTURE'S METHOD 



formalin and sometimes other sub- 

 stances such as osmic acid. The silver 

 is selective tending to impregnate a few- 

 cells completely which become black- 

 ened when it is reduced. E.xcept for the 

 occasional demonstration of the Golgi 

 Apparatus these methods do not reveal 

 details of the inner structure of nerve 

 cells like Neurofibrils and Nissl Bodies. 

 They are of great service in the demon- 

 stration of many non-nervous tissue 

 components, the parietal cells of the 

 stomach, bile canaliculi of the liver, 

 Rouget or perivascular cells, etc. 

 Golgi Method, Quick. For brains of new- 

 born animals, and of those 1 day to 30 

 days old. (Dr. J. L. O'Leary, personal 

 communication.) It is essential to 

 determine the age of the animal at which 

 the cell or fiber selected for study is 

 reaching maturity. For example, if 

 new born kittens are chosen, and the 

 area striata is the object of study, the 

 best impregnations of entering fibers are 

 obtained at 12 to 15 days after birth; 

 of short axon cells, at 18 to 21 da,ys; 

 and of pyramids at 21 to 24 days. 



Cut slices of brains 3-4 mm. in thick- 

 ness by quick cuts of a sharp scissors. 

 Fix in: potassium bichromate, 10 gm.; 

 osmic acid, 1 gm.; aq. dest., 330 cc. 

 Time of fixation must be determined 

 for each part of the CNS studied. In 

 general the older the animal, the longer 

 it is. After fixation, blot blocks of 

 tissue on filter paper and transfer to a 

 bottle containing f% aq. silver nitrate. 

 After 24 hrs. the reaction is complete. 

 Imbed in celloidin. Subsequent treat- 

 ment is very important. Place block in 

 95% ale. for about 5 min., remove and 

 blot dry. Place block on paraffin disc 

 mounted on a block holder in the orienta- 

 tion desired for cutting. Using a hot 

 teasing needle, melt paraffin around the 

 block so as to fasten block to paraffin. 

 Be sure that melted paraffin does not 

 creep up on the block. Use knife at 45° 

 angle to the block. Cut serially 80-100^. 

 Place each section as cut in order in 95% 

 ale. using Petri dishes. Be sure not to 

 miss first and last section of the block 

 for these are often more valuable than 

 the entire remainder of the block. 

 Using a spatula, transfer to another 95% 

 ale. after 5 min. After another 5 min. 

 transfer to oil of cloves, arranging in 

 serial order, by placing each section as 

 it enters oil of cloves near the edge of 

 the Petri dish so that it adheres to the 

 edge. When all sections are transferred 

 the group will be placed around the 

 circumference of the Petri dish. As the 

 sections start to retract from the edge, 

 begin to arrange them in the usual order 

 for serial sections. After clearing (clove 



oil 5 to 10 min.) transfer in serial order 

 to slides. Blot off excess of clove oil 

 and apply xylol, blot off xylol similarly 

 and apply a thin layer of Damar, using 

 the drop method. Let the slide dry on 

 an even surface adding more Damar as 

 necessary to keep sections protected. 



Gomori's Method. Silver impregnation of 

 reticulum, Gomori, G., Am. J. Path., 

 1937, 13,993-1001. Treat deparaffinized 

 sections of formalin fixed material with 

 0.5-1% aq. potassium permanganate. 

 1-2 min. Rinse in tap water ana 

 decolorize in 1-3% aq. potassium meta- 

 bisulphite, 1 min. Wash for several 

 minutes in running tap water. 2% 

 aq. iron ammonium sulphate (violet 

 crystals), 1 min. Wash in tap water 

 few minutes and then pass through 2 

 changes aq. dest. Impregnate for 1 

 min. in following solution : To 10% 

 aq. silver nitrate add | to | of its volume 

 of 10% aq. potassium hydroxide. While 

 shaking add strong ammonia drop by 

 drop until ppt. is completely dissolved. 

 Add carefully silver solution drop by 

 drop as long as resulting ppt. easily 

 disappears on shaking. Finally add 

 equal vol. aq. dest. Can be kept 2 days 

 in stoppered bottle. Rinse in aq. dest., 

 5-10 sec. Reduce in commercial forma- 

 lin diluted 5-10 times with tap water. 

 Wash under tap few min. Tone in 

 0.1-0.2% aq. gold chloride, 10 min. 

 1-3% aq. potassium metasulphite for 1 

 min. Fix in 1-2% aq. sodium thio- 

 sulphate (hyposulphite) for 1 min. 

 Wash under tap, dehydrate, clear 

 and mount. Reticulum black. Note 

 author's figures of sarcomata (Revised 

 by G. Gomori May 7, 1946). See Phos- 

 phatase. 



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, 0.69 gr.; 



