1. PURPOSE: To prepare the H & E stain (Harris) as per standard text literature & manufacture’s guidelines.

2. PRINCIPLE AND METHOD OF PROCEDURE USED FOR EXAMINATION: NA

3. PERFORMANCE CHARACTERISTICS: NA

4. TYPE OF SAMPLE: NA

5. PATIENT PREPARATION: NA

6. TYPE OF CONTAINER AND ADDITIVES: Conical flask

7. REQUIRED EQUIPMENT AND REAGENTS: Hematoxylin, absolute alcohol, potassium alum (aluminum potassium sulphate), distilled water, mercuric oxide/ sodium iodate glacial acetic acid.

8. ENVIRONMENTAL AND SAFETY CONTROLS: Universal safety precautions

9. CALIBRATION PROCEDURE (METROLOGICAL TRACEBILITY): The new batch of stains is used simultaneously with the in used batch of stains to determine the staining quality. For weighing of REAGENT/material calibrated weighing balance is used.

10. PROCEDURE(STEPS): H&E STAIN: • Dissolve the 2.5 gm of hematoxylin in the 50 ml of absolute alcohol and 50 gm of alum in the Luke warm 450 ml of DW.

• Mix the two solutions in a boiling flask and bring rapidly to boil.

• Add the 1.5 gm of mercuric oxide slowly after removing burner and then cool immediately by immersing the flask in cold water.

• The solution should assume a dark purple color on addition of the mercuric oxide.

• Add 10 ml of glacial acetic acid before use as this gives more precise and selective nuclear staining.

• Transfer to a suitable storage bottle.

11. QUALITY CONTROL PROCEDURE: Staining from every new lot (batch) of the REAGENTs (stains) should be done with the in use stain.

12. INTERFERENCE: Precipitate formation on storage. Discard the stains if the staining quality of sections poor.

13. PRINCIPLE OF PROCEDURE FOR CALCULATING RESULTS INCLUDING, WHERE RELEVENT, THE MEASUREMENT OF UNCERTAINITY OF MEASURED QUANTITY VALUES: NA

14. BIOLOGICAL REFERENCE INTERVALS OR CLINICAL DECISION VALUES: NA

15. REPORTABLE INTERVAL OF EXAMINATION RESULTS: NA

16. INSTRUCTION FOR DETERMINING QUANTITATIVE RESULTS WHEN RESULTS IS NOT WITHIN THE MEASUREMENT INTERVAL: NA

17. ALERT /CRITICAL VALUE, WHERE APPROPRIATE: NA

18. LABORATORY CLINICAL INTERPRETATION: NA

19. POTENTIAL SOURCE OF VARIATION: NA

20. REFERENCE: Histopathological laboratory techniques by Bancroft, 6th edition