568 METHODS AND FORMULAS MS 32.1-MS 32.2 



method: [15 yu paraffin sections of F 0000.4000 MacFarland and Davenport 1941 mate- 

 rial] —> water —>■ A, 1-2 days, 60°C. — > wash -* B, 1-2 days, 37°C. -^ quick rinse -^ 



C, till differentiated, 7-15 sees. —>■ wash, 1 min. — > D, 3-5 mins. -^ wash -^ E, 5-10 

 mins. — + wash, H niin. -^ /^, till sufficiently darkened, 15-30 sees. —* balsam, via 

 usual reagents 



RECOMMENDED FOR: nerve fibers in adrenal glands of mammals. 



32.1 Rogoff 1946 20540b, 21:59 



REAGENTS REQUIRED: A. 1% protein silver in a jar on the bottom of which has been 

 placed 4-6 Gm. clean metallic copper per 100 ml.; B. AMS 21.1 Bodian 1936; C. AMS 

 22.1 Bodian 1936; D. 2% oxahc acid; E. 5% sodium thiosulfate 

 method: [paraffin sections of mosquito larvae fixed in F 4000.0040 Petrunkewitsch 1933] 

 — * water -^ A, 36 hrs., 37°C. — > wash -^ B, 5-10 mins. — > wash -^ C, 2-5 mins. — > 

 quick rinse -^ D, 2-5 mins. -^ wash —>■ E, 5-10 mins. — * wash — * balsam, via usual 

 reagents 

 recommended for: cephalic ganglia of mosquitos. 



32.1 Silver 1942 20540b, 17:123 



reagents required: A. 0.2% protein silver; B. AMS 22.1 Silver 1942 

 method: [frozen, celloidin or paraffin sections of formaldehyde-fixed material, loose or on 



slide, but with embedding material removed] -^ water — » equal parts A and B, 45°C., 



till stained, 2-3 hrs. —* wash -^ balsam, via usual reagents 

 recommended for: nuclei, fine fibers, nerve terminations. 

 note : this method may be adapted to myelin sheaths by mordanting tissues for 1 week 



in 3% potassium dichromate 



32.1 Stage 1936 20540b, 11:155 



reagents required: A. 10% protein silver; B. 0.5% gold chloride; C. AMS 21.1 Stage 



1936; D. 5% sodium thiosulfate 

 method: [2 mm. slices fresh brain tissue] —> A, 2-20 days -^ 70% ale, rinse—' [cel- 

 loidin, via graded ales, and ale. -ether] -^^ [section 10-20 ju]^80% ale. -^ B, till 

 gray white, about 1 min. —y wash, 15 sees., or till differentiated — > dissolve celloidin 

 — » balsam 

 note: sections should not be exposed to alcohol lower than 85% between A and B. 



32.1 Ungewitter 1943 20540b, 18:183 



REAGENTS REQUIRED: A. 25% cliloral hydrate in 50% ale; B. \% protein silver in jar 

 containing 2-3 Gm. metallic copper per 100 ml.; C. AMS 21.1 Ungewitter 1943; 



D. 1% silver nitrate 



method: [fresh tissue] — > A, 24 hrs. — * [paraffin sections] -^ water — > B, 24 hrs. -^ rinse 

 — > C, 5-10 mins. — * wash — > D, 10-20 mins. — ^ wash — > [repeat C -^ D cycle till 

 sufficiently stained] — ^ balsam, via usual reagents 



32.2 Othek Protein Silver Methods 



32.2 Dawson and Barnett 1944 see MS 32.1 Davenport, Windle, and Rhines 1947 



32.2 Dublin 1943 Tech. Bull., 4:127 



REAGENTS REQUIRED: A. 1% protein silver; B.\% hydroquinone; C 0.5% gold chloride; 



D. 5% oxalic acid; E. 10% sodium thiosulfate 

 method: [paraffin sections of formaldehyde-fixed material] —* water —» A, overnight, 



37.5°C. -^ rinse -^ B, 10 mins. ~> rinse — > C, 5 mins. -* rinse -^ D, 5 mins. — > rinse 



—* E, 5 mins. — > wash -^ balsam, via usual reagents 

 RECOMMENDED FOR: demonstration of melanin. 



32.2 Moskowitz 1950 20540b, 25:17 



REAGENTS REQUIRED: A. 0.5% potassium permanganate; B. 5% oxalic acid; C. 1% pro- 

 tein silver; D. water 100, sodium sulfite 5, hydroquinone 1; E. 0.2% gold chloride; 

 F. 2% oxalic acid; G. 5% sodium thiosulfate 

 method: [smears fixed in F 3000.0000 Schaudin 1893 and well washed] -^ A,2 mins. -^ 

 wash -^ B, 2 mins. -^ thorough wash — > C, 36 hrs. at 35°C. — > rinse -^ F, 3 mins. —>■ 

 wash -^ G, 8 mins. — * thorough wash — > balsam, via usual reagents 

 RECOMMENDED FOR: extra-iiuclcar structures in protozoa. 



