RE:Retic stain

From:Charles.Embrey@carle.com

I always liked the Gordon- Sweet method better We always had very consistent
results.

2. CONTROL: cirrhotic liver

3. SILVER SOLUTION:
Add ammonium hydroxide, drop by drop, to 5ml of 5% silver nitrate until brown
precipitate clears.
Add 5ml of 3% sodium hydroxide ( precipitate forms )
Add ammonium hydroxide again, drop by drop, until precipitate just clears.
QS to 50ml with distilled water.

4. PROCEDURE:

4.1.   Deparaffinize, hydrate slides to distilled water.

4.2.   Treat with acidified potassium permanganate for 30 seconds.

4.3.   Place in 1% oxalic acid until pale - about 1 minute, wash well in
distilled water

4.4.   Mordant in 2.5% iron alum for 30 minutes, wash well with distilled
water.

4.5.   Treat with silver solution 1 minute and rinse in distilled water.

4.6.   Develop in 10% formalin for 1 minute, wash well.

4.7.   Tone in 0.2% gold chloride for 1 minute, wash well.

4.8.   Fix in 2.5% sodium thiosulfate 1 minute, wash well.

4.9.   Counterstain in nuclear fast red for 5 minutes.

4.10.  Wash well in water, dehydrate, clear and mount.

5. RESULTS:
Reticulum fibers......................................sharp black
Nuclei and cytoplasm..............................red to pink

Hope this helps.  On any retic you want to make sure that your ammonium
hydroxide is fresh or you may get spotty inconsistent results.

Charles R. Embrey Jr.  HT(ASCP), PA(AAPA)
Pathology Supervisor
Carle Clinic
Urbana, IL



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