4.Tissue processing

1.PURPOSE OF EXAMINATION For processing the biopsy tissues in a manner so as to ensure optimal results.

2.PRINCIPLE AND METHOD OF PROCEDURE USED FOR EXAMINATION As per instructions & operating manual of automated tissue processor manufacture.

3.STAFF RESPONSIBLE Senior technician

4.PERFORMANCE CHARACTERISTICS NA

5.TYPE OF SAMPLE Tissue bits (after grossing).

6.PATIENT PREAPARATION NA

7.TYPE OF CONTAINER AND ADDITIVES Perforated cassettes

8.REQUIRED EQUIPMENT AND REAGENTS Equipment: automatic tissue processor. Reagent: Alcohol (ascending grades), Xylene, Paraffin.

9.ENVIRONMENTAL AND SAFETY CONTROLS Universal safety precaution

10.CALIBRATION PROCEDURE (METROLOGICAL TRACEBILITY) calibration done once in a year through EQDC.

11.PROCEDURE (STEPS) By automated tissue processor

  1. 10% formalin -5 to 6 hrs
  2. 50% alcohol – 1hr
  3. 60% alcohol – 1hr
  4. 70% alcohol – 1hr
  5. 80% alcohol – 1hr
  6. 90% alcohol – 1hr
  7. 100% absolute alcohol -1hr
  8. Xylene 1-1 ½ hr
  9. Xylene 2-1 ½ hr
  10. Paraffin wax 1 -1hr
  11. Paraffin wax 2-1hr

Manual Tissue Processing

  1. Take 2 glass beaker of 500 ml capacity and add 70% acetone in first one and 80% of acetone in second beaker. put tissue cassette in first beaker for 30 minute
  2. After 30 minute remove tissue bucket from first bucket and put in second bucket for 30minute.
  3. Replace acetone of first bucket with 90% acetone.
  4. After 30 minute remove tissue bucket from second bucket and put in first bucket of 90% acetone for 30minute.
  5. Replace acetone of first bucket with 100% acetone.
  6. After 30 minute remove tissue bucket from first bucket and put in second bucket of 100% acetone for 30minute.
  7. After these steps tissue cassettes are kept in xylene for two changes each for 1 hour.
  8. Then cassettes are transferred to wax bath of histokinet.
  9. Wax bath is connected with electric power; wait till temperature reach at 65 C. put bucket inside for one hour.
  10. After 1 ½ hour remove cassettes from it add new wax in wax bath and again keep cassettes for 1 ½ hour.
  11. Tissue is ready for embedding.

12.QUALITY CONTROL PROCEDURE NA

13.INTERFERENCE NA

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