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Part
II: Microscopy
diagram 2 |
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Ziehl-Neelsen
staining
procedure
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Place
the numbered slides on a staining rack in
batches (maximum 12). Ensure that slides do not
touch each other |
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Flood
entire slide with Ziehl-Neelsen carbolfuchsin,
which has been filtered prior to use, or cover each
slide with a piece of filter paper if unfiltered
carbolfuchsin is used |
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Heat
the slide slowly until it is steaming.
Do not boil. Maintain steaming for three to
five minutes by using low or intermittent
heat. In no case must the stain boil dry |
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If
filter paper strips have been used, remove
them with forceps. Rinse each slide individually
in a gentle stream of running water until
all free stain is washed away |
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Flood
the slide with the decolourising solution for a
maximum of three minutes |
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Rinse
the slide thoroughly with water. Drain excess
water from the slide |
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Flood
the slide with counterstain |
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Allow
the smear to counterstain for 60 seconds |
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Rinse
the slide thoroughly with water. Drain excess water
from the slide. Allow smear to air dry. Do
not blot |
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