Acid Fast Staining

I. Objectives
At the end of the laboratory Experiment, the student shall be able to:

 To determine if the sample has the sign of the chronic disease of tuberculosis.
 To learn how to execute the Ziehl–Neelsen staining process.
II. Method
The procedures of the Acid Fast Staining are as follows:
Disinfect laboratory workplace using Lysol and Paper towel. Prepare the bacterial smear
on a clean grease free slide in an aseptic way. Sterilize the wire loop and put a small drop of
water. After this, put the phlegm sample into the smear, dry and heat- fixed. Then a drop of
carbol fuchsin stain, then heat the stain until the vapor began to rise and avoiding not overheating
the stain, then set aside for about five (5) minutes and wash off with distilled water.
III. Results

Figure 1: The group 2 sample.

The group 2 observed that the sample emits blue color resulting in the negative presence
of mycobacterium. The sample has no indication of the tuberculosis.
IV. Interpretation
During the laboratory experiment, the group 2 found it difficult to determine the presence
of tuberculosis, because at first try the group put too much methylene blue. On the second try, we
have successfully determined the blue non-acid fast cells, on another reading, there had been
cases that we observed the blue non acid fast cells because of the factors of improper handling of
the sample, According to the journal of bacteriology by Yegian, D., & Porter, K. R. (1944), they
have stated that “preparations made without the mechanical action of spreading instruments
failed to show these atypical forms, and (2) the relative abundance of non-acid-fast forms in
smears depends upon the amount of manipulation with the spreading loop or spatula”.
V. Conclusion
We found out that the sample has emitted no signs of the disease tuberculosis, because
the color emitted was blue non-acid fast cells and therefore has no presence of mycobacterium.
In the end of the laboratory experiment that day, the group 2 had learned what method should be
use in determining the staining process that we will be using in determining the presence of
mycobacterium.
VI. Reference
Yegian, D., & Porter, K. R. (1944). Some artifacts encountered in stained preparations of tubercle bacilli:
I. Non-acid-fast forms arising from mechanical treatment. Journal of bacteriology, 48(1), 83.