EXPERIMENT 2:

CELL STAINING TECHNIQUES

(Lugols Solution)

4 Bio 3 | Lopamia, Malicse, Marfil,

Introduction:

Tetrahymena sp.
ciliated single-celled
protozoan
freshwater organism
that inhabits
streams, lakes, and
ponds
cells are large 40-50
um
cells are inexpensive
to grow
grow rapidly to high
density in a variety of
media and conditions

Source:
http://www.zoology.ubc.ca/courses/bio332/Lab
s/CiliateProject/tetrahymena/TETRAweb.htm

Tetrahymena sp.
possesses many
core processes
conserved across a
wide diversity of
eukaryotes
(including humans)
that are not found
in other singlecelled model
systems

Source:
http://www.zoology.ubc.ca/courses/bio332/Lab
s/CiliateProject/tetrahymena/TETRAweb.htm

Lugols Solution
first made in 1829, and is named after
Jean Guillaume Auguste Lugol
an aqueous solution of Iodine and
Potassium iodine with a precise ratio of
1:2
also referred to as Iodone-Potassium
solution or IKI

Lugols Solution
Applications:
for detection of starch,
for disinfection purposes,
for the staining of bacteria according to
Gram,
and as a fixative.
It is also used in various experiments to
observe how a cell membrane uses
osmosis and diffusion, and in Schiller's
test in order to diagnose cervical cancer.

Lugols Solution
As a fixative:
Lugols iodine is able to kill cells without
distorting the structure

Lugols Solution
Detection of Starch
Starch consists of amylose (20-30%) and
amylopectin (70-80%). The ratio of
amylase:amylopectin varies depending
on the natural source of the starch.
Amylose is formed by branched and unbranched chains of glucose monomers.
[I5] - ions of Lugols solution intercalate
into starch molecules. Intercalation of
penta-iodide results in different colors of
the complexes: amylase is stained in
blue, amylopectin is stained in violet red.

Objective:
To identify the parts of the cells that are
made visible with the stain (Lugols
solution) used in the experiment

Results and Discussion

Group 1

Group
2

Group 3

Group 4

Group 5

Group
6

Group 7

Group 8

Group 9

Group 10

Observation:
The Tetrahymena appeared completely
black/brown in the scanner objective, LPO,
HPO and OIO.

Conclusion:

Conclusion:
Since Lugols iodine is an indicator test for
the presence of starch, it reacts by turning
into a black or dark blue color. It also stops
all the cellular functions of the cell. Lugols
iodine killed the cell, however the stain
was able to enter the cell and show the
nuclei and glycogen vacuoles making the
cell appear completely brown in the
microscope.

The End

4 Bio 3 | Lopamia, Malicse, Marfil,