SECTION TWO 



NILE BLUE SULPHATE 

 For demonstrating fatty acids and neutral fats 



Solution required: 



To lOO ml. saturated aqueous Nile Blue sulphate add 

 0-5 ml. cone, sulphuric acid; then boil under a reflux con- 

 denser for two hours; allow to cool; then use as follows: 



Technique: 



1. Fix small pieces of tissue in 10% formalin. 



2. Frozen sections are stained for about ten minutes to half an 

 hour at 37° C. ; or overnight at room temperature. 



3. Differentiate in 2% acetic acid. 



4. Rinse in distilled water; mount in Farrant. 



Results: 



Free fatty acids: blue. Neutral fats: red. 



References : 



Cain, A. J. (i947)- 

 Smith, J. Lorrain (1907). 



NILE BLUE-PICRO FUCHSIN 



(After Murray-Drew) 



For bacteria and actinomyces in pathological tissues 



Solutions required: 



A. Picro fuchsin (Van Gieson). 



B. Nile Blue sulphate aqueous i%. 



Technique: 



1. FormaHn-fixed material is embedded in paraffin wax, or 

 frozen sections may be employed. 



2. Take sections down to distilled water in the usual manner 

 then stain in picro fuchsin for two or three minutes. 



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