AR Slide - Background overfluorescing(20752 bytes)

 

This smear is showing over fluorescence of the background of a slide stained with auramine-rhodamine.  The AFB in the smear are  fluorescing the same color as the debris and are blurry, making it difficult to make out the morphology.  Someone inexperienced in the  reading of fluorescent smears might mistake this as being all debris causing a false negative report to be sent.   Any of the following mistakes during staining can contribute to this occurance:

  1. Insufficient washing off of the auramine-rhodamine stain.
  2. Insufficient decolorization of the smear because the smear was made too thick.
  3. The slides are not decolorized with acid-alcohol for the required length of time.
  4. Insufficient decolorization due to the slide not being  flooded  with acid alcohol until all visible traces of auramine-rhodamine are gone.
  5. Insufficient quenching with Potassium Permanganate.

 

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