1. PURPOSE OF EXAMINATION: To lay down the procedure for staining the Tissue Sections in Histo-pathology section at NCHLS Surat.
2. STAFF RESPONSIBLE: Lab technician & all the authorized signatories of reports in histopathology section.
3. PRINCIPLE AND METHOD OF PROCEDURE USED FOR EXAMINATION: Hematoxylin is oxidized to hematin (in situ with Iodate) which in acidic conditions binds to lysine residues of nuclear histones by linkage via a metallic ion mordant (aluminum) giving it blue/black color. Eosin Y is acidic colorant which binds with protein in cytoplasm.
4. PERFORMANCE CHARACTERISTICS: NA
5. TYPE OF SAMPLE: Tissue sections.
6. PATIENT PREPARATION: NA
7. TYPE OF CONTAINER AND ADDITIVES: NA
8. REQUIRED EQUIPMENT AND REAGENT: Hematoxylin & Eosin stain, Methanol, acid alcohol, Tap water, Jars & assembly.
9. ENVIRONMENTAL AND SAFETY CONTROL: Universal safety precautions.
10. CALIBRATION PROCEDURE (METROLOGICAL TRACEBILITY): 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.
11. PROCEDURE (STEPS) :
Manual method:
• Dewax sections in xylene giving two changes for 15 minutes.
• Put the section in 85% alcohol for 2 minute.
• Put the section in 80% alcohol for 2 minute.
• Dry the section in air.
• Dip the section in water for 2 times
• Stain with Harris Hematoxylin for 5 minutes. • Wash well in tap water for five Minutes till sections are blue.
• Dip in 1% acid-alcohol: 1-2 dips.
• Wash in sodium bicarbonate for 2 minutes.
• Stain in 1% Eosin for 30 second.
• Wash in tap water for 1-2minutes.
• Dehydrate through ascending grade of Alcohol.
• Clear in Xylene and mount with DPX after drying.
• RESULTS:
12. QUALITY CONTROL PROCEDURE: Discard the stains if the staining quality of sections poor.
13. INTERFERENCE: Contaminated Alcohol or Xylene
14. PRINCIPLE OF PROCEDURE FOR CALCULATING RESULTS INCLUDING, WHERE RELEVENT, THE MEASUREMENT OF UNCERTAINITY OF MEASURED QUANTITY VALUES: NA
15. BIOLOGICAL REFERENCE INTERVALS OR CLINICAL DECISION VALUES: NA
16. REPORTABLE INTERVAL OF EXAMINATION RESULTS: NA
17. INSTRUCTION FOR DETERMINING QUANTITATIVE RESULTS WHEN RESULTS IS NOT WITHIN THE MEASUREMENT INTERVAL: NA
18. ALERT /CRITICAL VALUE, WHERE APPROPRIATE: NA
19. LABORATORY CLINICAL INTERPRETATION: NA
20. POTENTIAL SOURCE OF VARIATION: NA
21. REFERENCE: Histopathological laboratory techniques by Bancroft, 6th edition