Histotechnology Technical methods

Stain for Nucelic Acids

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1. Bring sections to water.

2. Rinse sections in M - HCL at room temperature 1 min.

3. Place sections in preheated M - HCL at 60°c (this hydrolysis time will vary and depends on the fixative. For formalin fixed tissues 8 mins hydrolysis is required.

4. Rinse sections in M - HCL at room temperature 1 min.

5. Transfer sections to Schiff's reagent 45 min - 1 hr.

6. Rinse sections in bisulphite solution x 3 washes - 2 mins each.

7. Rinse well in water.

8. Counter stain with 1% light green in 1% acetic acid 40 sec.

9. Dehydrate, clear . Mount sections in DPX


--DNA Magenta --Cytoplasm Green


M - Hydrochloric acid

--Hydrochloric acid (conc) 8.5ml

--Distilled water 91.5ml

Schiffs reagent

--See PAS technique

Bisulphite solution

--10% potassium metablsuphite 5ml

--M HCL 5ml

--Distilled water 90ml

NOTE: No responsibility is assumed by The University of Nottingham or the Queens Medical Centre NHS Trust for any injury and/or damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods, products, instructions or ideas contained in the material herein. It is the users responsibilty to ensure that all procedures are carried out according to appropriate Health and Safety requirements.

© Copyright 1997 University of Nottingham Medical School Division of Histopathology. This page was last built on Mon, May 5, 1997 with Frontier. Thanks for looking in. Comments to James.Lowe@nottingham.ac.uk