RETICULIN STAIN PROTOCOL

(1) DEPARAFFINIZE AND HYDRATE TO DISTILLED TO WATER.

(2) OXIDIZE IN 0.5% POTASSIUM PERMANGANATE 5 min.

(3) RINSE IN RUNNING TAP WATER  2 min.

(4) 2% POTASSIUM METABISULFITE 2 min.

(5) RINSE IN RUNNING TAP WATER 2 min.

(3) SENSITIZE IN 2% FERRIC AMMONIUM SULFATE 10 min.

(4) RINSE WELL IN DISTILLED WATER 1 min.

(5) AMMONIACAL SILVER NITRATE SOLUTION 5 min.

(6) RINSE QUICKLY IN 3 CHANGES OF DISTILLED WATER 30 sec.

(7) 20% UNBUFFERED FORMALIN 5 min.

(8) RINSE IN RUNNING TAP WATER 2 min.

(9) TONE IN 0.2% GOLD CHLORIDE 10 min.

(10) RINSE IN RUNNING TAP WATER 1 min.

(11) 2% POTASSIUM METABISULFITE 2 min.

(12) RINSE IN RUNNING TAP WATER 1 min.

(13) REMOVE EXCESS SILVER IN 2% SODIUM THIOSULFATE 2 min.

(14) RINSE IN RUNNING TAP WATER 1 min.

(15) DEHYDRATE, CLEAR AND COVERSLIP.

 

* Sections should turn dark brown to black immediately after applying the formalin solution. If this reaction does not occur start over using freshly made solutions and acid clean glassware.