Staining of Negri Bodies |
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Abstract: | The following procedure for staining Negri bodies in sections is based on methods previously described by MacNeal, by Haynes, and by Richter:Fixation: 1. Zenker's solution 4 hours at 37°C or Dominici's 3 hours. 2. 70% alcohol, 12 to 18 hours at room temperature. 3. 80% alcohol, about 5 to 6 hours. 4. 90% alcohol, about 4 to 6 hours. 5. Absolute alcohol about 16 hours. 6. Ether and absolute alcohol aa, about 8 hours. 7. 16 to 24 hours in the following mixture: celloidin 1 g., methyl salycilate 25 cc., abs. alcohol 25 cc., ether 25 cc. 8. Chloroform and paraffin, 2 to 3 hours. 10. Paraffin, 1 to 1 1/2 hours. 11. Embed. staining: 1. Cut sections 4 to 5 μ. 2. Bring section to water and cover with Lugol's iodine for 10 minutes. 3. Decolorize with a 2% sodium thiosulfate (hypo). 4. Wash thoroly with water. 5. Cover with a mixture of equal parts of 0.5% phloxine and 1% eosin Y (National Aniline brand) and leave for 15 minutes. 6. Wash with water and stain 2 to 5 minutes in 0.1% azure B (National Aniline). 7. Wash with 96% alcohol and decolorize in a mixture of 2 parts absolute alcohol with 1 part clove oil, ordinarily for not more than 1/2 to 1 minute. 8. Dehydrate rapidly, clear, and mount in Yucatan Elemi.
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