GOLGI APPARATUS 



146 



GOLGI METHOD, QUICK 



hrs. Counterstain, dehydrate, clear 

 and mount. 



It is of interest that all silver prepara- 

 tions depend upon the treatment of the 

 tissue by fixatives which contain the 

 salt of a heavy metal. Silver, M. L. 

 (Anat. Rec, 1942, 82, 507-529) has 

 pointed out that silver micelles are not 

 adsorbed on to the Golgi material un- 

 less the cells have been treated with 

 salts of heavy metals — hence the pres- 

 ence of metals like cobalt and ura- 

 nium in the fixatives used for silver 

 techniques. 



Stains: 



1. Baker's Sudan Black Method (Baker, 

 J. R., Quart. J. Micr. Sci., 1944, 85, 

 1-71). Fix small pieces of tissue in 

 formol calcium (formalin 40%, 10 cc, 

 calcium chloride, anhydrous 10% aq. 10 

 cc, aq. dest. 80 cc.) for 3 days, embed in 

 gelatine. Harden block in formalin-cal- 

 cium-cadmium (formalin 10 cc, calcium 

 chloride, anhydrous 10% aq. 10 cc, 

 cadmium chloride, 10% aq. 10 cc, aq. 

 dest. 70 cc.) and then wash 3-4 hrs. in 

 running water. Section on freezing 

 microtome at 15m. Sections when fixed 

 to the slide are placed in filtered for- 

 malin-calcium-cadmium solution until 

 ready for staining. Then wash slides 

 for 3 min. in running water, pass 

 through 50% and 70% ale to a sat. solu- 

 tion of Sudan black in 70% ale, leave for 

 7 min. Pass through 3 lots of 50% ale, 

 rinse in aq. dest., counterstain, mount 

 in glycerine jelly or Apathy's medium. 

 This method does not demonstrate the 

 Golgi apparatus as a network but as a 

 series of discrete bodies, a form which is 

 claimed by some authors to represent 

 more nearly the true form of the Golgi 

 apparatus in living cells. Many of the 

 vesicles which are demonstrated by 

 this method also stain with neutral red. 



2. Baker's Neutral Red Method (Baker, 

 J. R., Quart. J. Micr. Sci., 1944, 85, 

 1-71). This is a vital method. Cells 

 are separated by teasing in a salt mix- 

 ture (sodium Chloride, 0.7% aq. 100 

 cc, calcium chloride, anhydrous, 10% 

 aq., 0.2 cc). To 3 drops of solution 

 containing a suspension of cells add 3 

 drops of a solution made up of Neutral 

 red 0.1% aq., 1 cc. and sodium and 

 calcium chloride solution (as above) 

 9 cc. Final concentration of dye is 

 0.005%. Mix cell suspension with dj^e 

 by sucking up with pipette and pass out 

 again. Leave preparation (covered) 

 for 20 min. Put 2 drops of mixture on 

 the glass of a compressorium in such a 

 way as to include some air. Cover edge 

 of compressorium with soft paraffin 

 (e.g. Vaseline). Examine under im- 

 mersion lens. 



Golgi Cox Method for adult nervous system. 

 — Written by J. L. O'Leary, Dept. of 

 Neuropsychiatry, Washington Uni- 

 versity School of Medicine, St. Louis 

 10, Mo. May 8, 1950.— Fix pieces 3-6 

 mm. thick in following fluid : add 20 cc 

 5% aq. potassium bichromate to 20 cc 

 6% aq. mercuric chloride. Dilute 16 

 cc. 5% aq. potassium chromate with 40 

 cc. aq. dest. and add this to the first 

 two. Do not agitate but leave in 

 fixative until scum forms on surface, 

 usually after 1^-2 months. When im- 

 pregnation is nearly complete, wash 

 rapidly, dehydrate through graded alco- 

 hols and imbed in low viscosity celloidin 

 (see Celloidin Imbedding). Cut cel- 

 loidin sections serially at 80 to 120 

 microns. Arrange in serial order on 

 slides (80% alcohol). Blot sections 

 dry and cover immediately with 1% 

 celloidin. When somewhat dry, bring 

 slides with sections to water. The 

 sections on each slide may thereafter 

 be treated as a unit. Run sections 

 from water into a saturated solution 

 of sodium sulfite. They rapidly turn a 

 yellow gray. Wash over night and de- 

 hydrate through graded alcohols to ab- 

 solute. Coat with the following var- 

 nish, applying it repeatedly in thin even 

 layers, and allowing each to dry par- 

 tially before applying the next (san- 

 darac, 75 gm.; camphor, 15 gm.; 

 turpentine C.P., 30 cc; oil of lavender, 

 22.5 cc; abs. ale, 75 cc; add castor oil, 

 7 drops. Mixture dissolves very 

 slowly). Since sections are somewhat 

 opaque, the varnish must dry for several 

 days until abs. ale has evaporated. 



Golgi Methods. Fundamentally these are 

 different from both the Cajal and Biel- 

 chowsky techniques which were later 

 developments. They depend upon a 

 preliminary fixation in a potassium 

 bichromate solution often containing 

 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. Except 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. — Written by J. L. O'Leary, 

 Dept. of Neuropsychiatry, Washing- 

 ton University School of Medicine, St. 

 Louis 10, Mo. May 8, 1950.— It is es- 



