International University, Vietnam National University - HCMC

BIOLOGY LABORATORY

LAB REPORT
Course name: Biology

Course’s ID: BT155IU

Instructor: Quang Thien Nguyen

Group: Five (5) Section: Saturday Morning Date: 24/11/2018

Group members: 1. Đặng Gia Hoàng - BTBTIU17071

2. Phùng Vân Thủy - BTBCIU17033

3. Huỳnh Thị Thảo Nguyên - BTBTIU17095

Instructor’s comment:

1
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY

Contents

PRACTICAL 1: MICROSCOPY, CELL OBSERVATION & OSMOSIS....................3

I. PLANT CELLS AND ANIMAL CELLS OBSERVATION.............................................................................................3

II. OSMOSIS IN PLANT CELLS........................................................................................................................................ 4

PRACTICAL 2: ORGANIC COMPOSITIONS OF THE CELL..............................7

I. CARBOHYDRATE............................................................................................................................................................7

II. PROTEIN.......................................................................................................................................................................... 8

III. LIPIDS............................................................................................................................................................................10

PRACTICAL 3: PHOTOSYNTHESIS &TRANSPIRATION................................ 12

I. PHOTOSYNTHESIS......................................................................................................................................................12

II. TRANSPIRATION......................................................................................................................................................... 13

PRACTICAL 4: ENZYMES....................................................................................... 16
I. AMYLASE........................................................................................................................................................................16

II. PROTEASE....................................................................................................................................................................18

III. CATALASE....................................................................................................................................................................20

PRACTICAL 5: CELL DIVISIONS: MITOSIS & MEIOSIS................................. 22

I. MITOSIS.......................................................................................................................................................................... 22

II. MEIOSIS.........................................................................................................................................................................23

THE END............................................................................................................................................................................ 24

2
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY

PRACTICAL 1: MICROSCOPY, CELL

OBSERVATION & OSMOSIS
I. PLANT CELLS AND ANIMAL CELLS OBSERVATION

1/ Introduction

 Plant cells are eukaryotic cells appear in green plants. A plant cell has a nucleus and
several organelles bounded a cell membrane. It is noticeable that a plant cell has cell
wall and chloroplast. Cell wall helps cell stand stably, meanwhile chlorophyll in
chloroplast converts sunlight into energy and gives plant cells their green color.
 Similarly to plant cells, Animal cells are eukaryotic cells and each cell has a nucleus
and numerous organelles. After observing plant cells and animal cells through
microscope by different magnifications, we can see the cells visually and find analogy
between them.
2/ Procedure
 Plant cells – onion epidermis cells

 Step 1: Split a piece of onion leaf into half. Then separate the outermost part.
 Step 2: Minimize air bubbles under the layer when putting the epidermis layer on the
slide.
 Step 3: Drop Lugol solution and cover with a coverslip.
 Step 4: Use the microscope to observe your slide from the lowest (4x) lens to 10x, and
40x.
 Animal cells – human cheek cell epithelium
 Step 1: Use a toothpick to scrape the inside of your cheek.
 Step 2: Put your scrapings on a clean slide and drop Lugol solution.
 Step 3: Add a coverslip on the top.
 Step 4: Use the microscope to observe your slide from the lowest (4x) lens to 10x, and
40x.

3/ Results

3
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY
4/ Discussion
a) What is the function of Lugol solution in these experiments?
 Lugol’s solution (LS) was developed 1829 by the French physician Jean Guillaume
August Lugol, initially as a cure for tuberculosis. It is a solution of elemental iodine (5%)
and potassium iodide (KI, 10%) together with distilled water. It has been used as a
disinfectant, a reagent for starch detection in organic compounds, in histologic
preparations, in dental procedures and in diagnosis of cervical cell alterations.

 In these experiments, Lugol solution is used to dye the cells so that the cells get better
observation via the microscope.
b) What is the difference between plant cells and animal cells?

Animal Cell Plant Cell

Cell wall Absent Present (formed of cellulose)

Shape Round (irregular shape) Rectangular (fixed shape)
Vacuole One or more small vacuoles One, large central vacuole
(much smaller than plant cells). taking up to 90% of cell volume.
Centrioles Present in all animal cells Only present in lower plant
forms (e.g. chlamydomonas)
Chloroplast Absent Plant cells have chloroplasts to
make their own food.
Plastids Absent Present
Plasma Membrane Only cell membrane Cell wall and a cell membrane
Lysosomes Lysosomes occur in cytoplasm. Lysosomes usually not evident.

Cilia Present Most plant cells do not contain.

Food Storage Glycogen Starch

Nucleus At the center Near cell wall

II. OSMOSIS IN PLANT CELLS

1/ Introduction
 Diffusion is defined as the movement of molecules from the area of more solute
concentration to the less solute concentration.
 Osmosis (includes Hypertonic, Hypotonic and Isotonic) allows water molecules pass
through a selectively permeable membrane (such as Cellular membrane) from a solution

4
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY
with a lower solute concentration to a higher one. Thank to osmosis, the concentrations
on each side of membrane keep balance. The types of molecules that can pass through
or permeate the membrane without stinting are limited.

2/ Procedure
 Step 1: Rind a thin epidermis layer (purple side) of the Zebrina leaf.
 Step 2: Add a small drop of 0.85% NaCl on 2 prepared clean glass slides.
 Step 3: Put the layers into two above clean glass slides, add coverslips.
 Step 4: Use the high power lens (40x) to observe the plant cells which are located in the
region that is not too dense.
 Step 5: Add 2-3 drops of 5% NaCl to the edge of the first coverslip. Observe the
differences.
 Step 6: Do the same way with the second coverslip but replacing 5% NaCl by distilled
water. Observe the differences.

3/ Results

5
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY
4/ Discussion
a/ Explain the phenomenon.
If the extracellular fluid has lower osmolarity than the fluid inside the cell, it’s said to
be hypotonic—hypo means less than—to the cell, and the net flow of water will be into the
cell. When adding water into Zebrina pendula leaf, water potential outside the cell greater
than inside the cell, so the cell gains water, which means water moves into the cells. This
process is called becomes Hypotonic.
In the reverse case, if the extracellular fluid has a higher osmolarity than the cell’s cytoplasm,
it’s said to be hypertonic—hyper means greater than—to the cell, and water will move out
of the cell to the region of higher solute concentration. When adding the 5% NaCl into
Zebrina pendula leaf, water potential outside the cell lower than inside the cell, so the cell
loses water and becomes Hypertonic.
In an isotonic solution—iso means the same—the extracellular fluid has the same
osmolarity as the cell, and there will be no net movement of water into or out of the cell.
When adding the 0.85% NaCl into Zebrina pendula leaf, water potential is the same as the
cell, so there is no net movement of water across the cell membrane. This is called Isotonic
solution.
b/ When putting plant cells in concentrated NaCl, plasmolysis happened. When
putting animal cells in water, hemolysis occurred. What makes the phenomenon in
plant cells different from in animal cells?

“Cell wall” which makes the phenomenon in plant cells different from in animal cells.
When osmosis repeats many times and the plant cell absorbs water to the point
where it can absorb no more. The cell is now turgid and the cell wall prevents the cell
from bursting. Because animal cells do not have a cell wall, they can burst in dilute
solutions.

6
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY

PRACTICAL 2: ORGANIC COMPOSITIONS OF THE

CELL
I. CARBOHYDRATE

1/ Introduction

 Glucose is found in plants and is used in respiration or it is consumed as fuel in animals

and plants. Excess glucose will be stored as starch in plants and glycogen in liver and
muscles of animals.
 The most stable starch is coiled helix. In amylose, the core of helix is of the right
dimension to accommodate iodine, this complex is deep blue.
 However, the glycosidic bond can be wrecked by heat, enzymes… In this experiment,
we will observe the starch granules and demonstrate the hydrolyzing of starch by heat
and acid.

2/ Procedure
a/ Task 1:
 Step 1: Put the scratched potato on the slide and observe the starch granules.
 Step 2: Then adding Lugol solution and observe the change in color.
b/ Task 2:
 Step 1: Put one drop of rice starch suspension onto a slide. Add 1 drop of Lugol solution
and compare with original Lugol solution.
 Step 2: Add 5ml of starch suspension into a test tube, then add 5 drops of concentrated
HCl solution, mix well.
 Step 3: Take out 1 drop of starch – HCl mixture onto the slide and test the color with 1
drop of Lugol solution.
 Step 4: Place this test tube into the boiling water bath.
 Step 5: Every 5 minutes, take out 1 drop of hydrolysed starch-HCl mixture and add 1
drop Lugol solution until no color change. Observe the change in color.

3/ Results
*Observe the Starch Granules: Starch BEFORE and AFTER adding Lugol solution.

7
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY
*Chemical detection of Starch:

4/ Discussion
a) Explain the phenomenon when adding Lugol solution to potato starch granules?
When adding Lugol solution to starch granules, the iodine molecules will fit the coiled helix
structure of the starch, making the complex. This complex has the deep blue color.

b) Explain the different color in Starch – HCl mixture after time of boiling. Based on
the color of spot, why can we say that the structure of starch is affected by
temperature?
When boiling the solution, color of the spot becomes lighter until there is no change in color.
Base on the color of the spot, we can state that structure of the starch is affected by
temperature because acid and heat cause a hydrolysis reaction which can wreck the starch
structure.

II. PROTEIN
1/ Introduction
 Proteins are polymers made of monomers called amino acids. Amino acids are organic
molecules possessing both carboxyl and amino groups. They link together by peptide
bonds to form long chains (polypeptides or oligopeptides).
 The biuret test is a chemical test used for detecting the presence of protein or peptide
bonds. In this test, we add alkaline solution and metal ions such as Cu, Zn … into our
sample. In the strong basic environment, when peptide bonds are present, the metal
ions will form a coordination complex with 4 nitrogen atoms from peptide bonds. The
complex of metal ions and nitrogen atoms make the color of solution changes to violet.
This color range from violet to red is dependent on the number of peptide bonds in the
solution.

8
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY
2/ Procedure
 Step 1: Put 2 ml of egg white and fresh cow milk into 2 separated test tubes.
 Step 2: Add 2 ml of 10% NaOH solution into each test tube.
 Step 3: Add 4 drops of 0.5% CuSO4 solution into the test tubes.
 Step 4: Observe the color change.

3/ Results

COLOR OBSERVATION
Protein After 10% After 0.5%
Original Color
Solutions NaOH CuSO4
Egg
White viscous Dark violet Blue precipitate
Albumin
Fresh
Opalescent Pale violet Violet
Cow Milk

White egg and Milk

4/ Discussion
a) Explain the function of 10% NaOH and 0.5% CuSO4 in Biuret reaction?
- NaOH creates the base environment of the biuret reaction by having OH-
- CuSO4 provides Cu2+ in order to link with nitrogen atoms to make the biuret reaction
occur.
b) After adding 10% NaOH, the phenomenon in egg white is different from in cow
milk, why?

After adding NaOH, the phenomenon of eggs occurs clearly and it is easy to observe. The
structure of protein in egg white is easily affected in the base environment. Besides,
adding NaOH to milk creates 2 parts in the test tube: emulsion and watery liquid leads
which leads them to denature. Furthermore, while albumin has disulfide bonds, casein in
milk does not have disulfide bridges so that there is a difference in 2 phenomena.

9
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY
c) Why is the color intensity in egg white different from in cow milk?
Because proteins in egg look like glue but protein in milk have a hydrophilic side, so color of
this reaction in egg is deep purple, look like a precipitate while color of the solution of milk is
light purple.

III. LIPIDS
1/ Introduction
 Lipids are a family of organic compounds that are mostly insoluble in water. Composed
of fats and oils, lipids are molecules that yield high energy and have a chemical
composition mainly of carbon, hydrogen, and oxygen.
 Lipids are not soluble in water. They are non-polar and are thus soluble in nonpolar
environments like in choloroform but not soluble in polar environments like water.
 The three main types of lipids are triacylglycerols, phospholipids, and sterols.
 Lipids perform three primary biological functions within the body:
 Serve as structural components of cell membranes
 Energy storehouses
 Important signaling molecules.
 Examples: fats, oils, waxes, certain vitamins, hormones and most of the non-protein
membrane of cells.

2/ Procedure
Step 1: the peanut soaked in water is sliced as slim as possible.
Step 2: Put the peanut in a glass slides and stain it by Soudan III solution in 10-minute.
Step 3: Using 20% Ethanol to wash the slide.
Step 4: Add coverslip on after putting on drop of immersion oil to the sample.
Step 5: Use microscope (100x) to observe the lipid granules stained in peanut cells.

3/ Results
Under microscope, we can see many red-orange dots.
They are lipids after stained by Soudan III.

10
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY
4/ Discussion
a) Why is Soudan 3 used to detect lipid?

Soudan III is a fat-soluble dye used to color nonpolar substances like oils, fat, waxes...
Therefore, it stains lipids red. Using Sudan III can show the amount and the location of
lipids on the slide, which allows to observe in a clearer view.

b) Why do we have to wash the stained sample with 20% Ethanol before
observation under microscope?

We have to wash away the stained sample with 20% Ethanol in order to wash the
excessive Soudan III that blend with the stained-lipid. Therefore, we can observe
the stained lipid clearly under microscope.

11
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY

PRACTICAL 3: PHOTOSYNTHESIS &TRANSPIRATION

I. PHOTOSYNTHESIS
1. Introduction
 Photosynthesis is a process of converting sunlight into useable chemical energy from
carbon dioxide and water that can be later released to fuel the plants. Photosynthesis
maintains atmospheric oxygen levels and supplies all the organic compounds and most
of the energy necessary for life on earth.
 Photosynthesis happens in chloroplasts. In chloroplast, a chemical compound called
chlorophyll which is an important pigment in absorbing and transferring light energy.

2. Procedure
a) Examination of oxygen formation
 Step 1: Prepare a bucket of water.
 Step 2: Inside water, place inside the beaker 10 waterweed brands, funnel, test tube
respectively.
 Step 3: Put the beaker under the light.

b) Examination of starch formation

 Step 1: Prepare leaves by covering a middle half of these leaves for 2 weeks.
 Step 2: Remove the cover, put each leaf into a test tube and place in the boiling bath for
5 minutes.
 Step 3: Pour all water out of test tubes, pour ethanol 70% in and then place them in the
boiling bath until the green color disappears.
 Step 4: Take out the leaves and stretch them out on a Petri dish, add Lugol solution into
the leaves and observe the color of the leaves.

3. Results
a) Examination of oxygen formation

Duration (days) Level of water (mm)

0 0
2 1
4 1.75
6 2.5

12
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY
b) Examination of starch formation

Color observation

After 1-week
Leave sample Original color After boiling After Lugol
covering

Uncovered part Green Green Green Yellow brown

Covered part Green Pale Green Pale Green Yellow

4. Discussion
a) Explain the phenomenon in lowering level of water in experiment 1
 The gas (oxygen) released by water weeds pushes water out in the test tube, making
the phenomenon in lowering level of water.

b) What will happen if we get the burned match to meet O2, CO2, H2?
 The burned match meets the O2: oxygen intensely brightens the bud of burnt match.
 The burned match meets the CO2: carbon dioxide darkens the bud of burnt match.
 The burned match meets the H2: hydrogen slight brightens the bud of burnt match.

c) Explain why the color of 2 area (covered and uncovered) is different

The uncovered areas have the color darker than the covered area because uncovered
areas consist of starch inside which is generated by photosynthesis process. This
substance combines with iodine to make the complex have deep blue color. Meanwhile,
the covered area lack of the procedure  no starch  no deep blue color.

II. TRANSPIRATION

13
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY
1. Introduction

Transpiration is a process by which water enters from the root and is transported to other
parts to moisturize trees through xylem. Although the leaf surface usually have a cover
which is a cuticle layer to avoid desiccation, the leaves also contain pores called stomata to
allow gases to enter and leave the leaf. This is important for photosynthesis process,
because carbon dioxide and oxygen can enter and pass out of the leaf only through an
opening the stomata flanked by two guard cells. And while these gases are moving between
inside and outside of the leaf, a large amount of water is also lost by this way.
2. Procedure
 Step 1: Prepare a small piece of absorbent paper and soak it into 3% CoCl2 solution for
2 minutes and then dry it.
 Step 2: Place this dried piece on a sticking tape by using forceps.
 Step 3: Quickly stick this tape onto a leaf.
 Step 4: Check the color of paper frequently and notice the time needed for this color
change.
3. Results

Color observation
Original color White
Absorbent paper After 3% CoCl2 Pink
After drying Blue
0 min Blue
5 min Pale blue
Duration after 10 min Pale blue + pink
applying on leaf 15 min Pale blue + pink
20 min Pale blue + pink
30 min Pink

Moreover, we observed that the paper on upper surface of the leaf change the
color slow than the above surface => the upper surface has fewer stomata.

14
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY
4. Discussion

Different trees will have different level of water-out through transpiration. Based on
the character(s) of the leaf, we can tell the level high or low. What is that
character(s)? Explain your answer.

Based on these 3 following characters of the leaf, we can tell the level high or low:
 Number of stomata: the opening of stomata allows vapor leave the leaf. Thus, the more
presence of the stomata, the higher level water - out. Various trees have various
numbers of stomata on their leaves.
 Thickness of cuticle: thicker the cuticle layer on a leaf surface, the lower level water
out.
 Size of the leaf: a leaf with bigger surface area will have the higher level of water-out
than a leaf with smaller surface area.

15
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY

PRACTICAL 4: ENZYMES
I. AMYLASE

1. Introduction
 Amylaseare enzymes that catalyses the hydrolysis of starch into sugars. Amylase is
present in the saliva of humans and some other mammals, where it begins the chemical
process of digestion.
 Amylases are found mostly in animal saliva, pancreas, intestine,… and in sprouting
seeds.
 All amylases are glycoside hydrolases and act on α-1,4-glycosidic bonds.
 There are three types of amylase:
 -amylase acts on 1,4--D-glucan-glucan glycoside.
 -amylase acts on 1,4--D-glucan-malto glycoside linkage.
 -amylase acts on 1,4--D-glucoside and glucan 1,6--glycoside linkages.

2. Procedure
 Step 1: Prepare 8 test tubes, mark them as the table below.

Temperature 40C RT 500C 1000C

Marked tubes 4-S 4-E RT-S RT-E 50-S 50-E 100-S 100-E

Be clear that: “4”: place in ice, “50”: place in warm water, “100”: place in boiled water, “RT”:
place in room temperature
 Step 2: Add 2ml amylase suspensions into test tubes “E” and Put them in the
appropriate condition given in the table above for 15 minutes.
 Step 3: Add 4ml of starch suspension into all test tubes. Continue to keep all reaction for
10 minutes in the same condition.
 Step 4: After the time indicated, take tubes out and add 1-2 drops of Lugol solution into
each tube and 2ml of water into each “S” tube
 Step 5: Mix well and observe the color.

3. Results

16
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY

Temperature (oC) Marked tubes Color observation

Before Lugol After Lugol

4 4-S white Deep blue

4-E white Deep blue
RT RT-S white Deep blue
RT-E white white
50 50-S white Deep blue
50-E white white
100 100-S white Deep blue
100-E white Deep blue

From left to right: The solution in 40C, RT, 500C and 100 0C

4. Discussion

a/ Compare “S” with ‘’E” tubes in each condition and explain the phenomenon:

 At 40C: the color in both “S” and “E” tubes before and after adding Lugol is the same.
At 40C, the enzyme amylase is not active. Without its catalytic activity, hydrolysis does
not occur and starch is not broken down to form glucose monomers. Thus, when
adding Lugol into the “E” tube, the color is dark blue, which is similar to when adding
Lugol into the “S” tube.
 At RT: “S” tube is dark blue while “E” tube is opalescent with a little purple. At RT, the
enzyme amylase is active. Starch is hydrolyzed. So, when adding Lugol into the “E”
tube, the color is opalescent with a little purple.
 At 500C: “S” tube is dark blue-black while “E” tube is opalescent. At 500C, the enzyme
amylase is active. Starch is hydrolyzed completely. Moreover, at this temperature,
amylase activity seems to be at its greatest, Therefore, when adding Lugol into the “E”
tube, the color is opalescent.

17
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY
 At 1000C: S” tube is dark blue-black while “E” tube is purple transparent.
At 1000C, the enzyme amylase is still active. With its catalytic activity, hydrolysis still
occurs and starch is still broken down. But because that amylase is denatured and lose its
normal function at high temperatures, so when adding Lugol into the “E” tube, the colorist
a little purple transparent color.

b/ Compares all “S” tubes in all conditions and all “E” tubes in all conditions. Explain
the phenomenon.

In all conditions, the color in all “S” tubes are the same while the color in all “E” tubes is
different
 Amylase is heat sensitive, the change in reaction temperature causes its change of
activities. Thus, at 40C, RT, 500C and 1000C, the ability to function of amylase is
different.
 At 40C, amylase is not active and starch is not hydrolyzed.
 At RT and 500C, amylase is active and starch is hydrolyzed
 At 1000C, starch is hydrolyzed but it is unlike what we see at RT and 500C when the
enzyme activates. (color is lighter than 40C’ but deeper than RT and 500C’)
 The lighter the color is, the greater the amylase activity is so 500C is the best
temperature (of four different temperatures) for amylase activity.
c/ What is the optimal range of temperature for amylase activity?
Through the results, we consider that at 50oC enzyme amylase digest starch fastest in
the same time, so we can conclude that a temperature around 50oC is the optimal
temperature for amylase activity.

II. PROTEASE
1. Introduction
 Proteases are enzymes that break peptide bonds between amino acids of proteins. This
process is called peptide cleavage. The enzymes use a molecule of water for this and
are thus classified as hydrolases
 The mechanism used to cleave a peptide bond involves making an amino acid residue
that has the character of a polarized peptide bond or a water molecule nucleotide
nucleophilic so that it can attack the peptide carbonyl group.
 Proteases occur naturally in all organisms and constitute 1-5% of the gene content.
Peptidases can break either specific peptide bonds, depending on the amino acid
sequence of a protein, or break down a complete peptide to amino acid.

18
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY
2. Procedure
 Step 1: Prepare 2 test tubes and put 5ml protease extraction (pineapple juice) into each
tube.
 Step 2: One tube is put in boiling water for 15 minutes, and the other left at room
temperature.
 Step 3: Add a small piece of boiled white egg into each tube.
 Step 4: Cover tubes, shake slightly and incubate at room temperature.
 Step 5: After 2 days, pour out the liquid and observe the results.

3. Results

Temperature At the beginning After 2 days

RT Shape: normal Shape: many scale
Color: yellow solution Color: dark yellow

100 Shape:normal Shape: a little scale

Color:white solution Color: still white

4. Discussion
a) Compare “S” with “E” tubes in each condition and explain the phenomenon

 At room temperature and 50oC: “S” test tubes have deep blue color, “E” test tubes have
white color  Amylase is active at these temperatures.
 At 4oC and 100oC: “S” and “E” test tubes are deep blue  starch is remained 
Amylase is inactive at these temperatures.
→ At high and low temperature, amylase is inactive, amylase active strongly at room
temperature because the color of RT-E tube is lighter than 50-E tube.

19
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY
b) Compare all “S”, “E” tubes in all conditions. Explain.

 “S” test tubes: all of them change the color into deep blue after adding Lugol solution.
This is because the starch is not changed it combines with I2, making complex
substance having deep blue color
 “E” test tubes:
 At room temperature and 50oC: the color is white (not changed), because amylase
performs a reaction that cutting starch into smaller molecules. The reaction between I2
and starch is not occurred  the color is not changed.
 At 4oC and 100oC: the color is deep blue, because amylase is inactive, reaction does not
occur, making deep blue complex.

c) What is the optional range of temperature for amylase activity

From approximately 35 – 40oC.

III. CATALASE

1/ Introduction
 Catalase is an anti-oxidant enzyme which is produced naturally in plants, animals, and
microbial cells. This enzyme functions to convert peroxide into less toxic substances.
 The ability of catalase is to convert hydrogen peroxide (H2O2) into oxygen (O2) and water
(H2O) when the body is infected with high level of hydrogen peroxide.
 Catalase is stored in peroxisome in plant cell and mitochondria in animal cell. Catalase
belongs to the group of oxidoreductase.
2/ Procedure
 Step 1: Prepare 4 test tubes, mark them 4, RT, 50, 100.
 Step 2: Add 5ml of enzyme suspension to each tube.
 Step 3: Mark a line at 5cm above solution surface.
 Step 4: Put them at appropriate temperature for 10 minutes.
 Step 5: Add 5 drops of H2O2 into each tube and record the results.

3/ Results

Temperature (oC) Time recorded (seconds)

4 29
RT 52
50 40
100 Long duration

20
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY

40C RT 500C 1000C

4/ Discussion
a) Why is there different in time when bubbles columns reach th 5-cm line
between different conditions?

 The air bubbles level: room temperature test tube > 50oC test tube > 4oC test
tube > 100oC test tube.
 This is because catalase at room temperature works best, decomposing hydrogen
peroxide into water and oxygen.
 At 4oC and 100oC, catalase is inactivated, so these test tubes provide very small quantity
of bubbles.

b) What is the optimal range of temperature for catalase activity?

The optimal temperature of catalase is roughly 37oC.

21
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY

PRACTICAL 5: CELL DIVISIONS: MITOSIS &

MEIOSIS
I. MITOSIS

1. Introduction
 Mitosis is the step in the cell cycle that the newly duplicated DNA is separated, and two
new cells are formed.
 This process is important in single-celled eukaryotes, as it is the process of asexual
reproduction.
 In multi-celled eukaryotes, mitosis is how a single zygote can become an entire
organism.
 Mitosis has several distinct stages, or phases, that will be discussed below:

1. Prophase The nucleolus, a rounded structure, shrinks and disappears.

The end of prophase is marked by the beginning of the organization
of a group of fibers to form a spindle and the disintegration of the
nuclear membrane.
2. Metaphase The chromosomes, each of which is a double structure consisting of
duplicate chromatids.
Line up along the midline of the cell.
3. Anaphase Each chromatid pair separates into two identical chromosomes that
are pulled to opposite ends of the cell by the spindle fibers.
4.Telophase The chromosomes begin to decondense, the spindle breaks down,
and the nuclear membranes and nucleoli re-form.
The cytoplasm of the mother cell divides to form two daughter cells,
each containing the same number and kind of chromosomes as the
mother cell.

* The stage, or phase, after the completion of mitosis is called “interphase”.

22
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY
2. Results

Phase Pictures
1. Prophase

2. Metaphase

3. Anaphase

4. Telophase

II. MEIOSIS
1/ Introduction
Meiosis is the process by which sex cells (gametes) are made in the reproductive organs. It
involves the reduction division of a diploid germline cell into four genetically distinct haploid
nuclei.

The process of meiosis consists of two cellular divisions:

 The first meiotic division separates pairs of homologous chromosomes to have the
chromosome number (diploid  haploid).
 The second meiotic division separates sister chromatids (created by the replication of
DNA during interphase I).

Meiosis I Meiosis II

Prophase I Chromosomes condense, Prophase II Chromosomes condense, nuclear

nuclear membrane dissolves, membrane dissolves,
homologous chromosomes centrosomes move to opposite
form bivalents, crossing over poles (perpendicular to before).
occurs.
Spindle fibres from opposing Metaphase II Spindle fibres from opposing
Metaphase centrosomes connect to centrosomes attach to
I bivalents (at centromeres) and chromosomes (at centromere)
align them along the middle of and align them along the cell
the cell. equator.

23
 International University, Vietnam National University - HCMC
BIOLOGY LABORATORY
Anaphase Spindle fibres contract and split Anaphase II Spindle fibres contract and
I the bivalent, homologous separate the sister chromatids,
chromosomes move to chromatids (now called
opposite poles of the cell. chromosomes) move to opposite
poles.

Chromosomes decondense, Telophase II Chromosomes decondense,

Telophase nuclear membrane may reform, nuclear membrane reforms, cells
I cell divides (cytokinesis) to form divide (cytokinesis) to form four
two haploid daughter cells. haploid daughter cells.

3. Result

MEIOSIS I MEIOSIS II
Prophase I Prophase II

Metaphase I Metaphase II

Anaphase I Anaphase II

Telophase I Telophase II

THE END

24