GOLGI APPARATUS 



112 



GOLGI METHODS 



cc; 2% osmic acid, 10 cc, at room 

 temperature 1-3 hrs. Wash in running 

 water then in aq. dest., 5 hr. 2% osmic 

 acid at 25 °C., 12-16 days. Wash for 

 24 hrs. in running water, dehydrate, 

 clear in chloroform and imbed. 



6. Kolatchew's method, Nassonov, D. 

 N., Arch. f. Mikr. Anat., 1924, 103, 

 437-482. Fix in 3% aq. potassium 

 bichromate, 10 cc; 1% chromic acid, 

 10 cc. ; and 2% osmic acid, 5 cc, 24 hrs. 

 Wash in running water 24 hrs. 2% 

 osmic acid, 40 °C., 8 hrs. 3-5 days at 

 35°C. Wash in aq. dest., dehydrate, 

 clear, and imbed. 



7. Weigert's Mann-Kopsch method as 

 modified by Gatenby (Lee's Microt- 

 omist's Vade-mecum. Ed. 9. Ed. by 

 Gatenby, J. B., and Cowdry, E. V., 

 London, 1928) . Fix in Mann's corrosive 

 osmic acid sol. (sat. aq. corrosive sub- 

 limate in salt sol., 10 cc ; 1% osmic acid, 

 10 cc) j-3 hrs. or more. Wash in aq. 

 dest., 15-30 min. 2% osmic acid, room 

 temperature 10-14 days. Wash in run- 

 ning water 2 hrs. or more. Dehydrate, 

 clear, and imbed. In the sections 

 Gatenby was able to extract the black- 

 ening step by step with turpentine and 

 thus to considerably improve the prep- 

 arations. 



8. Ludford's modification, Ludford, 

 R. J., J. Roy. Micr. Soc, 1926, 107-109 

 has experimented at length with osmic 

 acid methods and states that his best 

 results have been obtained as follows. 

 Fix mammalian and avian tissues in 

 Mann's corrosive osmic sol. 18 hrs. 

 Wash in aq. dest., 30 min. 2% osmic 

 acid at 30 °C. for 3 days. Water at 

 30°C. 1 day, dehydrate, clear, imbed in 

 paraffin and section. A useful variant is 

 to fix in same way and wash in aq. dest. 

 Then osmicate at 35°C. for 3 days, first 

 day in 2% osmic, second in 1% and third 

 in 0.5%. Leave in water for 1 day at 

 35 °C. He recommends various counter- 

 stains. 



The writer prefers the Ludford tech- 

 niques. See Lee (pp. 313-326) for a 

 critical statement of the problem; also, 

 Owens, H. B. and Bensley, R. R., Anat. 

 Rec, 1929, 44, 79-109 for a careful study 

 of factors influencing the osmic acid 

 changes and for their ferric chloride 

 osmic method. Before placing any 

 reliance in the Golgi apparatus as an 

 indicator of cellular activity it is essen- 

 tial (1) to make sure that the technique 

 being used brings to light all the Golgi 

 apparatus, not only a part of it and (2) 

 not to mistake either mitochondria or 

 droplets formed from them for parts of 

 the Golgi apparatus particularly when 

 it is in a dispersed condition spread 

 about in the cytoplasm. But when 



every known precaution is taken the 

 surface and volume of this peculiar 

 structure can be measured quantita- 

 tively by means of a special technique 

 in spinal ganglion cells (Covell, W. P., 

 Anat. Rec, 35, 149, 1927). According 

 to Monne, L., Protoplasma, 1939, 42, 

 184-192 it can be demonstrated by 

 polarized light. Tarao, S., Cytologia, 

 1940, 11, 261-281 has described a novel 

 method, which involves the digestion of 

 frozen sections with trypsin and colora- 

 tion of the Golgi apparatus with Nile 

 blue sulphate, the value of which re- 

 mains to be determined. An account 

 of alterations in hepatomas is of interest 

 (Dalton, A. J. and Edwards, J. E., J. 

 Nat. Cancer Inst., 1942, 2, 565-575). 

 Localization of alkaline phosphatase in 

 Golgi region of cytoplasm (Deane, 

 H. W., and Dempsey, E. W., Anat. 

 Rec, 1945, 93, 401-417). 



Golgi Cox Method. For adult nervous 

 system (Dr. J. L. O'Leary, personal 

 communication). Fix pieces 3-6 mm. 

 thick in following fluid: add 20 cc. 5% 

 aq. potassium bichromate to 20 cc. 

 5% 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 verj^ 

 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 



