260 Cytoplasmic Elements (chap. 18) 



fixation: Mann's osmic sublimate: 18 hours. 



osmic acid, 1% (1 gm./lOO ml. water) 50.0 ml. 



mercuric chloride, saturated aqueous, plus 0.37 



gm. sodium chloride 50.0 ml. 



procedure: 



1. Wash blocks of tissue in distilled water: 30 minutes. 



2. Impregnate, 2% osmic acid: 3 days, 30°C 



2% osmic acid: 1 day, 35 °C 

 1% osmic acid: 1 day, 35 °C 

 0.5 7o osmic acid: 1 day, 35 °C 



3. Wash in distilled water: 1 day. 



4. Dehydrate, clear, and embed. 



5. Section 6-7 microns, mount and dry. 



6. Deparaffinize, clear, and mount. 



results: 



golgi apparatus — black 



yolk and fat — black (these may be bleached out with turpentine) 



comments: 



If it is advantageous to have mitochondria stained on the same slide, 

 follow deparaffinization (step 6) by hydrating to water (include cau- 

 tious treatment with 0.125% potassium permanganate), and stain by 

 Altmann Method (page 263). Mitochondria will be crimson. 



DaFano's Method (culling, 1957; cowdrv, 1952)^ 



fixation: 3-18 hours in: 



cobalt nitrate 1.0 gm. 



distilled water 100.0 ml. 



formalin 15.0 ml, 



solutions: 



Ramon y Cajal's developer: 



hydroquinone 2.0 gm. 



formalin 6.0 ml. 



distilled water 100.0 ml. 



sodium sulfite, anhydrous 0.15 gm. 



Gold chloride: 



gold chloride stock soliuion (page 410) 1.0 ml. 



distilled water 80-90.0 ml. 



