ABSTRACT

The aim of the first experiment which is the use of microscopy is to use the
microscope in a correct manner and also to examine the specimen sample under the
microscope. For the first experiment, two animal cell and two plant cell were used in the
experiment. The sample specimen chosen were chicken and fish for animal cell and onion
and leave for the plant cell. The specimen were prepared and put on the glass slide and is
covered using a cover slip. The specimen then is viewed under the light microscope using the
appropriate magnification. The main difference between a plant cell and an animal cell is
plant cell contains a rigid cell wall while animal cell does not. The second experiment is the
stain and identify the specimen given in the laboratory. Two specimen were used as sample
for this gram staining experiment which were E. coli and yeast. Both of the specimen is
transferred to the glass slide with an inoculation loop. The gram staining processes uses
crystal violet, methylene blue, iodine, decolourizer, and safranin. From the experiment it is
known that E. coli is gram negative because it was stained pink and yeast is gram positive
which after the gram staining experiment the colour purple .

1.0 OBJECTIVES
EXPERIMENT 1: THE USE OF MICROSCOPE
i.
To learn the correct method of using light microscope
ii.
To examine specimen samples under light microscope
EXPERIMENT 2: STAINING AND IDENTIFICATION OF CELL
i.
ii.

To choose suitable staining agent for inspection of different types of specimens

To prepare slide using Gram staining methods

2.0 LITERATURE REVIEW

A microscope is an optical instrument for viewing objects that are too small to be seen
by naked eye. The science of investigating small objects using such an instrument is called
microscopy. The term 'microscopic' means minute or very small, not visible with the eye
unless aided by a microscope. The most common microscope used in laboratories is the light
microscope. The light microscope has the magnification of 4x, 10x, and 40x.
The Gram stain is the most important staining procedure in microbiology. It is used to
differentiate between gram positive organisms and gram negative organisms. Hence, it is a
differential stain. Gram staining involves a four-part process, which includes crystal violet,
the primary stain iodine, and the mordant a decolourizer made of acetone and alcohol
safranin, the counterstain. Gram negative and gram positive organisms are distinguished from
each other by differences in their cell walls. These differences affect many aspects of the cell,
including the way the cell takes up and retains stains.
Gram-positive bacteria are bacteria that give a positive result in the Gram stain test.
Gram-positive bacteria take up the crystal stain used in the test, and then appear to be purplecoloured when seen through a microscope. This is because the thick peptidoglycan layer in
the bacterial cell wall retains the stain after it is washed away from the rest of the sample, in
the decolourization stage of the test.

Gram-negative bacteria cannot retain the violet stain after the decolourization step;
alcohol used in this stage degrades the outer membrane of gram-negative cells making the
cell wall more porous and incapable of retaining the crystal violet stain. Their peptidoglycan
layer is much thinner and sandwiched between an inner cell membrane and a bacterial outer
membrane, causing them to take up the counterstain safranin and appear pink.
3.0 HYPOTHESIS
Yeast bacteria is a gram positive while E. coli is a gram negative bacteria.
4.0 MATERIALS AND METHODOLOGIES
EXPERIMENT 1
1. Microscope
2. Glass slides
3. Cover slips
4. Scalpel/Razor blade
5. Tweezers
6. Specimen samples
Animal cell

Chicken
Fish

Plant cell

EXPERIMENT 2

Onion
Leaf

1. Bunsen burner
2. Microscope
3. Glass slide
4. Cover slips
5. Tweezers
6. Water bottle
7. Crystal violet solution (95% dye content)
8. Methylene blue solution (90% dye content)
9. Iodine solution
10. Gram decolourizer solution
11. Gram safranin solution
12. S. cerevisae
13. E.coli