344 Histochemistry (chap. 21) 



Immerse 3 minutes in 3 solutions of NaCl (0.85%, 1% and 2%) 

 before incubation. 



Burstone Method: 



Fix in cold acetone, 8-24 hours. Pieces 3 mm. thick. Double-embed 



(page 339). 



solutions: 



Riitenbiirg and Seligman Method: 

 Solution A: 



sodium 6-benzoyl-2-naphthyl phosphate^ 25.0 mg. 



distilled water 80.0 ml. 



acetate buffer (0.5M) pH 5.0 20.0 ml. 



make hypertonic by adding 2 gm. NaCl to each 100 ml. of solution. 



Solution B\ 



tetrazotized diorthoanisidine 50.0 mg. 



distilled water 50.0 ml. 



make alkaline with NaoCOs. 



Burstone Method: 



Either of two substrates may be used.^ 



a. Naphthol AS-MX phosphate 



b. 6-benzoyl-2-naphthyl phosphate 



Dissolve 5 mg. in 0.25 ml. DMF (N, N-dimethyl 



formamide)^ and add distilled water 25.0 ml. 



0.2IVI acetate buffer, pH 5.2 25.0 ml. 



diazotized 4-amino-2,5-diethoxybenzanilide, 



C.I. 37175 (fast blue BBN salt)^ 30.0 mg. 



manganese chloride, 10% 2 drops 



Shake and filter. 



procedure: 



Riitenburg and Seligman Method: 



1. Transfer frozen sections or cryostat sections to substrate (solution 

 A) room temperature: 1-2 hours. 



2. Wash, 3 changes: 10-5 minutes each, in cold saline (fresh tissues) 

 or water (fixed tissues). 



' Dajac Laboratories. 



* Dajac Laboratories. 



' Matheson, Coleman and Bell. 



•General DyestiifT Corporation. 



