Dr Piyush Tailor

1. PURPOSE OF EXAMINATION: To lay down the procedure for Iron stain in Histo-pathology section at NCHLS Surat.

2. PRINCIPLE AND METHOD OF PROCEDURE USED FOR EXAMINATION: The section is treated with dilute hydrochloric acid to release ferric ions from binding proteins. These ions then react with potassium ferrocyanide to produce an insoluble blue compound (the Prussian blue reaction).

3. STAFF RESPONSIBLE: lab. technician under supervision of resident doctor/tutor.

4. PERFORMANCE CHARACTERISTICS: NA

5. TYPE OF SAMPLE: Histopathology section slide.

6. PATIENTS PREPARATIONS: NA

7. TYPE OF CONTAINER AND ADDITIVES:

8. REQUIRED EQUIPMENTS AND REAGENTS:

EQUIPMENTS: Acid cleaned glasswares.

REAGENT: 2% potassium ferrocyanide, 2% HCL, Safranin, alcohol, xylene, DPX

9. POSITIVE CONTROL: Normal Bone Marrow Biopsy.

10. ENVIRONMENTAL AND SAFETY CONTROL: Universal safety precautions

11. CALIBRATION PROCEDURE: NA

12. PROCEDURE(STEPS):

• Take slide on which staining has to be performed along with positive control.

• Bring section to distilled water.

• Transfer section in to fresh solution of equal parts of 2% aqueous potassium ferrocyanide and 2% HCL for 30 minute.

• Wash thoroughly in several changes of distilled water.

• Counter stain with Safranin for 10-15 seconds.

• Wash in water, dehydrate with alcohol, clear in xylene and mount with DPX.

• Dehydrate in alcohol, clear in xylene, mount with DPX

Result

• Nuclei : Red

• Iron : Blue

• Cytoplasm : Pink

13. QUALITY CONTROL PROCEDURE: The newer batch of stain is tested before being put in to use by checking their efficiency with the batch of stain already in use.

14. INTERFERENCE: Inadequate quality/improper dilution

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

16. BIOLOGICAL REFERENCE INTERVALS OR CLINICAL DECISION VALUES:NA

17. REPORTABLE INTERVAL OF EXAMINATION RESULTS: NA

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

19. ALERT/ CRITICAL VALUE, WHERE APPROPRIATE: NA

20. LABORATORY CLINICAL INTERPRETATION: Interpret test result after true positivity of control slide.

21. POTENTIAL SOURCE OF VARIATION: NA

22. REFERENCE: Histopathological laboratory techniques by Culling’s.