PARAWORLD Training Resources

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F.A.S.T. AFB User Guide: Staining Procedures for Acid-Fast Bacilli (AFB) Detection
F.A.S.T. Products used in specimen staining: One (1) Bottle F.A.S.T. Auramine O AFB Stain One (1) Bottle F.A.S.T. Decolorizer/Quencher

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1. Process Sample
Process and prepare sample fully, as seen in the F.A.S.T. AFB User Guide: Slide Preparation. Allow slide to completely cool before staining.
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2. Apply Stain
Use 8-10 drops of F.A.S.T. Auramine O Stain, or until smear is covered. Let stand for approximately 1 minute.
11 10 12

3. Rinse
Gently rinse slide with water. Be careful to avoid washing specimen off of slide. Carefully shake slide to remove excess water. Note: Tap water may contain chlorine, which can interfere with fluorescence. Avoid using if possible.

Minute
7 6 5

1 2

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4. Apply Decolorizer
Use 8-10 drops of F.A.S.T. Decolorizer/Quencher, or until smear is covered. Let stand for approximately 1 minute.
11 10 12

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5. Rinse
Rinse slide again with water.

Minute
7 6 5

1 2

6. Dry Slide
Allow slide to air dry. If necessary, gently blot excess water with a lint free tissue.

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7. Examine
Examine the slide using the ParaLens AdvanceTM LED fluorescence microscope attachment, or other fluorescent microscope. See reverse side for examples of AFB stained with F.A.S.T. reagents.

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F.A.S.T. AFB User Guide: Staining Procedures

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F.A.S.T. AFB Guide: Slide Review

Image 1 Image 2

Mycobacterium tuberculosis, viewed at 400x magnification. A dark background helps to clearly observe the fluorescence of the AFB. Image 3

The clump of bacteria in the center is magnified at greater detail where single bacilli can be seen even clearer. (Original photo taken at 400x, digitally modified) Image 4

Mycobacterial bacilli sometimes fluoresce at varying degrees, as seen in this picture. Note that there are single bacilli as well as clumped bacilli here. (400x)

AFB appearing as a brilliant green against an orange/black background (possibly due to residual stain). While the AFB are clear, it is best to wash the slides completely so that the background remains a contrasting black color. (400x)

Note: For instructions on detecting AFB using SureFocusTM Microscope Slides, please consult the SureFocus Microscope Slide User Guide, also available from QBC Diagnostics.

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Copyright QBC Diagnostics Inc, 2009-2011. All rights reserved. QBC, QBC Diagnostics, F.A.S.T., ParaWorld, ParaLens, ParaLens Advance, and SureFocus are trademarks of QBC Diagnostics.

Form No. 260 Rev C