=====7.SOP FOR H & E STAINING PREPARATION=====

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