Activity No.

2
Direct Fecal Smear

John Carlo G. Villanueva BSMT2-Sanguis A 02/21/21

Objectives:

At the end of this activity, the students are expected to:

1. state the advantages and disadvantages of direct fecal smear;
2. discuss the principle and purpose of direct fecal smear

Direct Fecal Smear (Saline and Iodine)

The direct fecal smear (or direct wet mount) is most useful for the detection of trophic forms
amebae and flagellates, allowing the observer to study the motility of the organisms, which is
often characteristic. Wet films are particularly appropriate for immediate examination of
bloody mucus and other samples recovered during endoscopic examination of the lower
intestinal tract, and for other fresh specimens in which protozoal disease is suspected.

In DFS, about 2 mg feces (amount forming a low cone at the tip of the applicator stick) are
emulsified in a drop of 0.85% NSS on a glass slide. A cover slip is placed and the sample is
examined under a microscope using a low power microscopic lens.

This is a routine method of stool examination primarily useful in the detection of motile
protozoan trophozoites. In this preparation, the trophozoites appear very pale and transparent.
Trophozoite can be stained to demenstarte nuclear morphology using Nair’s buffered
methylene blue (BMB). Entamoeba cystoplasm will stain pale blue and the nucleus, darker blue.

Protozoan cysts can also be seen in a direct fecal smear. A weak iodine solution can be used as
a temporary stain to demonstrate nuclei. Alternatively, a new mount can be prepared with
iodine alone. The cytoplasm will stain golden yellow/yellow, the nucleus will be pale refractile,
and the glycogen will be deep brown. Helminth eggs and larvae can also be detected uring this
preparation.

Direct fecal smear is of value only when the infection is quite heavy. It may not detect the
parasite in light infections.
Materials and Reagents:
Clean glass slides
Coverslips
Markers/glass pencil for labelling
Wooden applicator sticks
Normal saline solution (0.85%)
Lugol’s iodine solution (1%)

Procedure:

1. With the use of a marker or glass pencil, label the glass slide with patient’s name or
identification number on the left side of the slide.
2. Place a drop of saline on one slide and a drop of Lugol’s iodine on the other end.
3. With an applicator stick, pick up a small portion of the fecal sample about 2mg, and spread it
evenly on the saline in circular motion. Using the same applicator stick, pick up from another
portion of the sample and spread it in the same manner on the iodine drop.
4. Dispose the used applicator stick in a disinfectant.
5. Place a cover slip in each drop by holding it in an angle position, one side touching the smear
and lowering it slowly to avoid formation of bubbles.
6. Scan smear first under LPO covering the entire area of the coverslip.

https://youtu.be/uxRerKqT73g
 Post – Laboratory Questions

1. During the preparation of a fecal smear, it is important to prevent the formation of air
bubbles. What is the reason for this? (3pts)
- Air bubbles should be avoided because it can cause you to have a problem in distinguishing
bubbles from the real specimen. Bubbles impede species' free movement, such as ciliates. At the
point where the air enters the solution that was put in the smear, the bubbles cause optical
artifacts. It appears that the air bubble is enclosed by a dark ring. Any areas of the specimen are
hidden by this dark ring, and it makes observation more difficult.

2.A drop of iodine is also used in preparation of the fecal smear. What is the purpose of iodine.
(2pts)
- In wet mounts of concentrated fecal content, an iodine solution is also used. It is useful for
staining glycogen in protozoan cysts and making nuclei visible. The purpose of using iodine
solution is to stain the other structures of a parasite that cannot be seen on a Normal Saline
Solution Wet Mount.