1. What structures are visible in the unstained Tetrahymena sp.? With iodine?

With

methyl green?

In an unstained Tetrahymena sp. fast- dot like organisms are observed

under the scanner. These are transparent structures with colorless outlines and

may be used for identification of some of the organelles such as vacuoles and cell

membrane.

With the cell stained with iodine, the nuclei and the glycogen vacuoles are

now visible as the stain enters the cell and locomotory organelles such as cilia can

also be seen.

With the cell stained with methyl green, macronuclei and micronuclei can

be observed as well as its nuclei.

Figure 1. Unstained Tetrahymena sp. under the scanner

Figure 2. Vacuoles and cell membrane of unstained Tetrahymena sp. under LPO

Figure 3.Tetrahymena sp. stained with Lugol’s Iodine (IKI)

Figure 4.Tetrahymena sp. stained with Methyl green

 Figure 5.Tetrahymena sp. stained with Methyl green observed under HPO

2. Do all stains enter the cell or just render more contrast to the background?

All stains entered the cell. With the cell stained with iodine, it rendered a

contrast to the background because it gave off a darker color. With the cell stained

with methyl green, it blended in with the background displaying the cell’s outline

and the visible structures.

3. By providing a dark background, would the cell structures look clearer?

Dark background enhances the contrast of the image to bring out fine

details. Reduction in intensity of the light that had not passed through the

specimen would create a grey background and increase contrast even more, with

some parts of the specimen darker and other parts of the specimen brighter than

the background.
4. Tabulate the different structures seen under each of the stains used. Explain the
mechanism for each reaction.

OBSERVATION MECHANISM/REACTION

Iodine is used in chemistry as an indicator

for starch. When starch is mixed with
Locomotory iodine in solution, an intensely dark blue
organelles such as color develops, representing a starch/iodine
Tetrahymena sp. cilia are already complex.
stained with visible and ass the Starch is a substance common to most
Lugol’s Iodine stain enters the cell, plant cells and so a weak iodine solution
the nuclei and the will stain This polysaccharide is produced
glycogen vacuoles by all green plants as an energy store. One
are now visible. chain contains hundred to thousand
glucose units, which form a helical
structure in which iodine can be trapped
which results in the blue color of starch
upon iodine staining.

Methyl green has seven methyl groups

Tetrahymena sp. Macronuclei and
Stained with micronuclei of the
Methyl green cell can be observed
and the nuclei can
also be seen.
rather than crystal violet's six. This seventh
group is easily lost and the dye reverts to
crystal violet.
For that reason there is invariably a
quantity of crystal violet mixed with the
methyl green. It is also a basic dye used as
a chromatin stain, as a differential stain for
RNA and DNA, and as a tracking dye for
DNA in electrophoresis. It stains cell
nuclei light green. This superior
formulation of methyl green is suitable for
use with a wide range of enzyme
substrates.