MODULE 6 Troubleshooting Smear Preparation and Staining Interactive Exercise

1. 32 MODULE 6Troubleshooting Smear Preparation and Staining Interactive Exercise

2. 33 Smear Preparation

3. 34

4. 35

5. 36

6. 37

7. 38 Question: Identify the Problem?

8. 39 Staining

9. 40

10. 41 What Are the Causes of Pale Staining AFB?

11. 42

12. 43 Causes of Pale Staining AFB Concentration of CF <0.3%: Staining time less than 5 minutes Inadequate heating of carbol fuchsin on the slide Excessive exposure time with an acid alcohol decoloriser may remove the carbol fuchsin from the AFB

13. 44 Time Effect of Carbol Fuchsin on Intensity of AFB

14. 45

15. 46 What Are the Causes of Excessive Counterstaining ?

16. 47 What Are the Causes of Excessive Counterstaining ? Leaving counterstain on slide for longer than one minute Methylene blue concentration > 0.3%

17. 48

18. 49

19. 50

20. 51

21. 52

22. 53

23. 54 What Are the Causes of Insufficient Decolorisation?

24. 55

25. 56

26. 57

27. 58 Troubleshooting and Miscellaneous Excessive heat-fixing of smear Glass scratches and AFB Soot on underside of smear

28. 59

29. 60

30. 61

31. 62 Summary Errors in smear preparation and staining contribute to false-negative and false-positive results Every step of the protocol needs to be performed to your best possible level