LESSON 1.

MICROSCOPE

1. MICROSCOPE
1.1. Introduction
A microscope is an instrument that magnifies an image of an object so that it is visible to the human
eye.
Use the following formula to determine the total magnification of a specimen.
Total Magnification = Ocular lens Magnification x Objective lens Magnification
1.2. Light Microscope Components
1.2.1. Optical components include Ocular (eyepiece), an Objective lens, a Condenser, and an
Illuminator (light source: lamp or mirror).
- Ocular: An eyepiece, or ocular lens, is a type of lens that is attached to a variety of
microscopes. It is named because it is usually the lens that is closest to the eye when
someone looks through the device. The ocular lens is normally embroidered with its
magnification (5x, 6x, 10x, or 15x)
- Objective lens: The magnification (or power) of each objective lens is engraved on the
side of the objective. Objective lenses come in various magnification powers, with the most
common being 4x, 10x, 40x, and 100x.
1.2.2. Mechanical parts: Focus knobs, Mechanical stage

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1.3. Using
To protect the microscope and slide, please follow the instructions in the following order:
- Plug in the power, and turn on the switch. Look into the eyepiece to adjust the light
source.
- To begin, observe the specimen with a low-magnification objective lens (4x).
- Put the slide on the stage, hold it on by the stage clip, and adjust it to the center of the
light source.
- Observe the slide, turn the coarse adjustment knob gently until the tip of the objective
lens nearly touches the slide.
- Then turn the coarse adjustment knob until the specimen is visible (if not clearly, adjust
the fine adjustment knob gently until it is visible).
- For greater magnification, a larger objective lens (10x, 40x) can be used.
1.4. Microscope maintenance
- The microscope must be stored in a cool, dry place. Cover carefully to prevent dust
from sticking to the objective lens and eyepiece.

2. SAMPLE PREPARATION
- Add a drop of water or a drop of glycerine to the slide.
- Put the specimen in a drop of water or glycerine.
- Cover the cover slip on the slide (figure). Lower the cover slip gently to avoid the
formation of air bubbles (Place a cover slip at a 45–degree angle to the slide and gently
let it fall on top of the slide).
- Observe with objective lenses 4x, 10x, and 40x

3. PRACTICE
3.1. Materials: purple onion; potato; yeast
3.2. Chemicals: Lugol’s solution; NaCl 8%; methylene blue
3.3. Practice:
3.3.1. Animal cells: oral epithelial cells
- Use a bamboo toothpick to gently scrape the oral mucosa (the cells from the inside of
your cheek).
- Dip the tip of the toothpick into the drop of Lugol’s solution on the slide.
- Cover the slide and observe under the microscope at the 40x objective.
3.3.2. Plant cells: onion cells and contraction of the protoplasm (plasmolysis)
- Peel the onion, use a razor knife to separate a few pieces of the purple onion's cuticle,
and soak them in the watch glass containing water.
- Select several thin strips placed on the slide. Cover with a cover slip and observe under
the microscope at 10x and 40x objectives.

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 - Plasmolysis: Use tissue paper to absorb the water under the cover slip, and add 1-2
drops of 8% NaCl to the edge of the cover slip. Observe what happens to the cuticle
fragment.
- Deplasmolysis: Use tissue paper to absorb (remove) NaCl solution under the cover slip,
and drop 1-2 drops of distilled water on one edge of the cover slip. Observe what
happens to the cuticle fragment.
3.3.3. Microbial cells: yeast
- Put a drop of methylene blue on the slide. Add a drop of yeast broth to the drop of
methylene blue. Cover with a cover slip and observe under the microscope at the 40x
objective.
3.3.4. Starch granules: potato starch granules
- Use a sharp needle to scrape the potato slices
Put some (very little) of this powder on the slide in a drop of water (prevent the starch
granules from overlapping)
- Cover the slide with a cover slip and observe under the microscope at 10x and 40x
objectives.
- Turn the fine adjustment knob slightly to see the concentric circle in the starch granules.
4. REPORT
4.1. Draw pictures and fully annotate the parts of the microscope. Describe the function of each
part.
4.2. Draw pictures (take photos), annotate, and describe the characteristics of each type of cell.
Note: it is necessary to note the components of the cell (walls, plasma membrane, cytoplasm,
nucleus).
4.3. Draw pictures (take photos), annotate, and describe the characteristics of starch granules.

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 LESSON 2. CELL MEMBRANE
1. INTRODUCTION
All organisms are formed by cells. Each cell is the unit structure and function of the
organism. The plant cell is surrounded by a cellulose wall. The cell wall maintains the cell’s shape
and protects the cell from mechanical damage. The cell membrane is the layer between the cell
wall and the cytosol. Cytosol contains organelles. Each organelle plays a different role in the cell.
Cells, as well as their organelles, have a lipoprotein membrane that separates them from their
surroundings. The intact cell membrane exhibits selective permeability. It retains the necessary
organic and mineral metabolites; controls its metabolism with the environment; maintains its
osmotic pressure; and allows the movement of water across the membrane by osmosis.
Every factor that affects the integrity of the membrane structure affects the above-mentioned
function of the cell.
2. Practice
2.1. Materials
Red beetroot
2.2. Chemicals
Absolute alcohol.
2.3. Practice
Effects of temperature and organic solvent on the permeability of cell membranes
- Cut the beetroot into 7 equal-sized pieces with a cork borer. The length of each sample is about
4 cm. Ensure that no peel is left on the samples.
- Put the beetroot pieces into a beaker, rinse them under running water for a few minutes to remove
all pigments from the broken cells (stop when the wash water is free of pigment). Dry the
beetroot pieces with tissue paper.
- Heat treatment:
+ Label 7 test tubes from 1 to 7, and write your group number on the surface of the test
tubes.
+ Put a piece of beetroot into each test tube.
+ Put 5 test tubes (numbers 2, 3, 4, 5, 6) in water at 5 different temperatures (40, 50, 70,
100, and - 10C) for 10 minutes. After heat treatment, put all the tubes in a test tube rack.
- Tube 1 - 6: add 15 ml of distilled water/tube.
- Tube 7: add 15 ml of absolute alcohol (cover the tube with a piece of nylon).
- Keep test tubes at room temperature, and wait for 15 minutes. Use a forceps to remove the
beetroots from the tubes, shake the tubes gently, and observe the color intensity of the solutions.
Compare the color intensity of solutions in tubes with that in the standard tube.
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Note:
+ The sample size must be THE SAME.
+ Heat treatment time must be THE SAME.
+ The time to soak the samples in test tubes must be THE SAME.
3. REPORT
Record the results in a table. Evaluate the color intensity in each test tube by the number
of plus signs (+). Make comments on the effect of high/ low temperature and absolute
alcohol on the permeability of cell membranes. Explain.

Tube 1 2 3 4 5 6 7
Color
intensity

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 LESSON 3. THE CHEMICAL COMPONENTS OF CELL
1. INTRODUCTION
Cells contain large amounts of water and organic components such as carbohydrates, lipids,
proteins, nucleic acids, and minerals.
1.1. Carbohydrates
- Starch, glycogen: is the storage form of carbohydrates (starch in plants, glycogen in animals).
They can be recognized by Lugol's reagent, which gives a blue-violet color to starch or red to
glycogen (for glycogen requires several other properties to distinguish it from dextrin, an
intermediate product of starch hydrolysis), e.g. turbidity, formation of sediments, etc.
These carbohydrates will be hydrolyzed into simple sugars (monosaccharides) to provide for the
metabolic activities of the cell, for example when the seed germinates.
- Reducing sugars: the appearance of simple (reducing) sugars will be recognized by Fehling's
solution: when heated in the alkaline environment of Fehling's solution, monosaccharides bearing
C = O radicals in the structure will reduce Cu2+ to Cu+, producing red (Cu2O) or yellow (CuOH)
sediment.
1.2. Lipids
Lipids are present in cells in many forms: triglycerides (fats, oils), phospholipids, glycolipids, and
steroids. Most easily observed are the oil droplets in the cells of the storage tissues of plants or oil-
containing seeds (coconut, peanut). Lipids were stained orange by the Soudan reagent.
1.3. The protein
Amino acids are polymerized by peptide bonds to form proteins. Proteins can be identified by a
variety of reagents because they react color in the presence of amino acids or cyclic amino acids.
Proteins or compounds containing 2 or more peptide groups (-CONH-) in a strongly alkaline
medium will form with Cu2+ a complex called biuret-Cu with a pink-violet color (Biuret reaction).
2. Experiment
2.1. Materials
Potatoes, soaked mung beans, mung bean sprouts, soaked peanuts, soaked white beans, fresh milk,
egg whites.
2.2. Chemicals
Fehling Reagent, Soudan III Reagent
Lugol’s reagent,
CuSO4 5%, NaOH 30%
2.3. Practice
2.3.1. Starch
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Prepare 2 test tubes:
- Tube 1: Crush 1 potato sample with 10 mL of distilled water, and remove residue with a filter
cloth. The filtrate is added to the test tube.
- Tube 2: Contains 10 mL of distilled water.
Add 1 drop of Lugol to each test tube. Shaken. Observe and record the phenomenon.
2.3.2. Reducing sugar
- Sugar extract: Crush 20 mung bean sprouts in a mortar. Add 20 mL of water, and mix well. Allow
to settle for 10 minutes, filter with a filter cloth, and discard residue. Put the filtrate into a beaker.
- Rinse this filter cloth with water to make sure that there is no sugar left in the filter cloth. Do the
same with 20 mung beans soaked in water for 1 hour.
- Reducing sugar test: Prepare 3 test tubes as information in the table. Use a marker pen to write
the numbers 1, 2, 3, and your group number on the surface of the tube
Use a pipette to get the exact volume
Test tube Fehling Reagent Water Extract of bean Extract of soaked
(mL) (mL) sprouts mung beans
1 (Control) 3 3 - -
2 3 - 3 -
3 3 - - 3

- Heat treatment: All 3 tubes were left in the boiling water for 5 min. Observe the phenomena
occurring in the test tubes.
2.3.3. Lipids
Prepare 3 test tubes:
- Tube 1: grind 5 peanuts in 10 ml of water. Filter out the residue. Collect the solution through the
filter into the test tube.
- Tube 2: contains 3 ml of cooking oil (use a dropper to get cooking oil, do not use a pipette
because it is very difficult to clean).
- Tube 3: contains 3 ml of distilled water.
Add 5 drops of Soudan III to each test tube. Observe the phenomenon in all 3 test tubes.
2.3.4. The protein
2.3.4.1. Plants:
Place 1 thick slice of soaked white beans on the slide (the thickness of the sample is about 1 mm).
Add 2 drops of CuSO4 and cover with a cover slip. After 10 min, unload the cover slip, and add 1
drop of NaOH. Observe the color appearing on the slice.

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2.3.4.2. Animal:
Prepare 3 test tubes:
- Tube 1: contains 5 mL of egg white solution.
- Tube 2: contains 5 mL of milk.
- Tube 3: contains 5 mL of distilled water.
Add 5 drops of CuSO4 to each test tube. Shake. Add 2 drops of NaOH, and shake gently. Observe
the color change.
3. REPORT
Describe the phenomenon that occurred in all experiments. Explain why the phenomena happen
(based on color reactions with reagents).

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 LESSON 4. ENZYME
1. INTRODUCTION
Enzymes catalyze all reactions in living cells. The main function of enzymes is to speed up
reactions. Enzymes are proteins. As a result, some factors, such as high temperatures, strong acids
or alkalis, organic solvents, and heavy metals can denature proteins, causing enzymes to lose their
activity.
The rate of the reactions depends on substrate concentration, enzyme concentration,
temperature, pH, etc.
Amylase in germinated seeds involves the hydrolysis of starch to glucose
(C6H10O5)n + nH2O nC6H12O6
To evaluate the activity of amylase, Lugol’s solution is normally used. The color of Lugol
will turn a dark blue if starch is available.
2. PRACTICE
2.1. Material
Germinated mung beans.
2.2. Chemicals
- Starch solution 0,2 %
- Lugol’s solution
- Buffer solution pH 3, 4, 5, 6, 7, 8
2.3. Practice
Effect of some factors on amylase activity:
2.3.1. Amylase extraction
Grind about 20 germinated mung beans. Add 20 ml of distilled water, and crush the beans
with a pestle. Allow it to settle for 10 minutes. Filter through a filter cloth to obtain the filtrates
(containing amylase).
2.3.2. Effect of temperature on amylase activity
Prepare 4 test tubes labeled 1, 2, 3, and 4. Write your group number on the surface of the test tubes.
Add 1 mL of starch solution to each test tube (use a pipette to get the exact volume of the solution).
Heat treatment:
- Tube 1: keep in ice water (about 5C).
- Tube 2: keep at room temperature (about 30oC).
- Tube 3: keep in 50C water
- Tube 4: keep in boiling water (100oC).
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 After 10 minutes, shake the filtrate, and add 1 mL of the filtrate to each tube.
Keep the same temperature conditions for 15 minutes
Put all the tubes in a test tube rack (note: put tube 4 into a cup of water to cool it down).
Arrange all the tubes on a stand and add one drop of Lugol’s solution to each tube to evaluate
the remaining starch in the tubes.
Note:
Lugol’s solution is added to each tube AT THE SAME TIME.
Shake well, and IMMEDIATELY observe the phenomenon.
2.3.2. Effect of pH on amylase activity
Prepare 6 test tubes labeled 1, 2, 3, 4, 5, and 6. Add to each test tube:
- 1 ml of starch solution
- 1 ml of buffer solution (pH 3, 4, 5, 6, 7, 8)
- 2 ml of the filtrate
Keep at room temperature for 7 minutes. After that, add one drop of Lugol’s solution to each
tube to evaluate the remaining starch in the tubes.
Note:
Lugol’s solution is added to each tube AT THE SAME TIME.
Shake well, and IMMEDIATELY observe the phenomenon.
3. REPORT
3.1. Evaluate the color intensity in each test tube by the number of plus signs (+).
Explain the effects of different temperatures on amylase activity.
Estimate the optimal temperature for amylase activity.

Temperature (oC) 5 30 50 100

Color intensity

3.2. Evaluate the color intensity in each test tube by the number of plus signs (+).
Explain the effects of different pH on amylase activity.
Estimate the optimum pH for amylase activity.

pH 3 4 5 6 7 8

Color intensity

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 LESSON 5. CELLULAR RESPIRATION
1. INTRODUCTION
Cellular respiration occurs in most cells of both plants and animals. Cells must
continuously obtain energy from the environment and use it to make adenosine triphosphate
(ATP).
The two important pathways to extract energy from nutrients include aerobic
respiration and anaerobic respiration (fermentation).
1.1. Aerobic respiration
C6H12O6 + 6O2 6CO2 + 6H2O + ATP
Substances in respiration include carbohydrates, proteins, lipids, or organic acids.
The release of CO2 in respiration is demonstrated through the ability of KOH to absorb
CO2 to form a BaCO3 precipitate.
1.2. Anaerobic respiration (fermentation)
The summary reaction of alcoholic fermentation
C6H12O6 → 2 C2H5OH + 2 CO2
2. PRACTICE
2.1. Material
Carrot
Potato
Germinated mung beans
Yeast
2.2. Chemical

Methylene blue 0,1 % H2O2 1 %

Resin gaiac 2 % Cooking oil
Saturated Ba(OH)2 solution Saccharose 30 %

2.3. Practice
2.3.1. Enzyme detection
2.3.1.1.Dehydrogenase
Put in the first test tube ( 30 mm) 2- 3 mm thick carrot slides, avoiding overlap. Place
methylene blue solution in the test tube that is about 1 cm above the carrot slices. Pour a layer of
cooking oil about 0.5cm onto the methylene blue surface.
The second test tube does the same but does not have carrot slices (control tube).
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 Keep two test tubes at room temperature for 60 minutes.
Observe the phenomenon. Compare the difference between the two tubes.
2.3.1.2.Oxidase
Cut two thin slices of potato (across the bud). Add one drop of resin guaiac to the first
potato slide. Observe the phenomenon.
Put the second slice of potato in boiling water for 5 minutes. Add one drop of resin guaiac
to it. Observe the phenomenon.
2.3.1.3.Catalase
Grind 10 g of potatoes in a mortar. Add 20 ml of distilled water and crush the potatoes with
a pestle. Filter through a filter cloth to obtain the filtrate. Distribute the filtrate evenly between 2
test tubes ( 18 mm).
Keep the first test tube at room temperature.
Place the second test tube in boiling water for 10 minutes, then transfer it to a beaker
containing water for rapid cooling.
Add 2 ml of H2O2 to each test tube. Observe the phenomenon.
2.3.2 . Aerobic respiration:
Put in a flask a handful of germinated mung beans.
Cover the flask with a rubber stopper with a U-tube and a funnel
(Figure 1). Seal the other end of the U-tube and funnel with wet cotton so that
CO2 does not escape. Wait for 90 minutes.
After that, remove the wet cotton and quickly immerse the glass tube
tip in a test tube containing Ba(OH)2. For faster observation, water can be
poured into the flask through a funnel to push CO2 from the flask to the test
tube. Figure 1

Observe and explain the phenomenon.

2.3.3. Anaerobic respiration:
Prepare 3 test tubes (Figure 2)
- Tube 1: 5 mL of saccharose solution and 5 mL of distilled water. Picture 2
Figure 2
- Tube 2: 5 mL of saccharose solution and 5 mL of yeast suspension
- Tube 3: 5 mL of distilled water and 5 mL of yeast suspension.
Use rubber balloons to seal the test tubes. Observe the phenomenon after 90 minutes.
3. REPORT
Observe the experiment, describe, draw/ take photos, and explain the phenomenon.

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 LESSON 6. PHOTOSYNTHESIS
1. INTRODUCTION
1.1. Paper chromatography
In chromatography, there is a mobile phase and a stationary phase.
The stationary phase is the phase that doesn't move and the mobile phase is the phase that does
move. The mobile phase moves through the stationary phase picking up the compounds to be
tested.
In paper chromatography, the mobile phase is the solvent. The stationary phase in paper
chromatography is the strip or piece of paper that is placed in the solvent. This paper
chromatography uses capillary action to move the solvent through the stationary phase.
The retention factor, Rf, is a quantitative indication of how far a
particular compound travels in a particular solvent. If the Rf
value for the unknown compound is close to or the same as the
Rf value for the known compound then the two compounds are
most likely similar or identical.
Rf = distance the solute (d) moves divided by the distance
traveled by the solvent front (D)
Figure 1. Rf
Rf = d / D
1.2. The Photosynthesis equation
6CO2 + 6H2O → C6H12O6 + 6O2
2. PRACTICE
2.1. Materials
Fresh leaves
Leaves have been partially shaded for 2 days
Aquatic plant
2.2. Chemical compounds
Acetone, benzene, Petroleum ether, Whatman filter paper, paper chromatography, saturated
Ba(OH)2 solution, ethanol 70%, Lugol’s solution
2.3. Practice
2.3.1. Analysis of pigment composition of leaves by chromatographic method
a. Extraction of leaves
- Grind some leaves in a clean and dry mortar. Add 20 ml of acetone and crush the leaves with a
pestle. Filter through Filter Paper, and collect the leaf extract into a clean, dry test tube.

Figure 2
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Use a nylon bag and a rubber band to seal the test tube.
Observe and note the color of the leaf extract.
b. Chromatography:
- Preparing chromatographic paper:
+ Size of chromatographic paper: 10 x 10 cm
+ Use a pencil to draw the starting line and the ending line as shown in Figure 3. Roll the paper
into a tube, and hold it with 2 staples at both ends of the tube, the 2 edges of the paper do not
overlap (Figure 3).

Figure 3
+ Pour the leaf extract (pigment solution) into a small petri dish.
+ Dip the tip of chromatographic paper in the leaf extract. When the pigment solution reaches the
start line (pencil line), remove the chromatographic paper, and dry it with a hair dryer.
+ When the pigment stain is completely dry, dip the tip of the paper into the pigment solution
again, wait until the pigment solution touches the start line, take it out and dry again. Repeat these
steps about 4 times (Figure 4)

Figure 4
Preparing the mobile phase (in a fume hood).
+ Prepare a clean and dry graduated cylinder.
+ Use a pipette to take 3 ml of benzene and put it in the cylinder. Add 27 mL of ether to 3 mL of
benzene to form the chromatographic solution (add ether into the cylinder up to 30 ml). Use a
nylon bag and a rubber band to seal the cylinder.
Process of chromatography
+ The solvent will be added to the Petri dish. Put the chromatographic paper into the Petri dish
(The level of the chromatographic solution should be a few mm below the starting line). The petri
dish and chromatographic paper of all groups will be placed together in a sealed box.
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+ When the mobile phase reaches the end line, remove the chromatographic paper and dry it (with
a hair dryer). Determine the Rf value.

Figure 5
2.3.2. Photosynthesis releases oxygen gas
Turn a funnel upside down over some aquatic plant in a
basin of water. This funnel will be covered by a test tube
filled with FULL water (Figure 6).
Lay out this system under sunlight. Then, the air bubbles
will be released from the cut of the aquatic plant. The gas
produced by this plant will be collected in the test tube.
After 1h, cover the mouth of the test tube with your finger,
turn it upside down, and bring a near-dead match to the
mouth of the test tube.
Figure 6. Photosynthesis
Take note of the phenomenon.

2.3.3. Photosynthesis using CO2

- 2 glass bottles numbered 1, 2
- Blow air from the mouth into 2 bottles: bottle 1 and bottle 2, then seal with a rubber cap.
+ Bottle 1: Use a syringe to inject 10 mL of saturated Ba(OH)2 solution. Shake.
+ Bottle 2: Use a syringe to inject 10 mL of distilled water. Shake.
Take note of the phenomenon.
- Wash both bottles.
- Put in bottle number 2 a fresh branch. Blow into both bottles: bottle 1, and bottle 2, and then
close the cap.
- Layout both bottles under a strong light source. After 90 minutes, use a syringe to inject 10 mL
of saturated Ba(OH)2 solution into each bottle. Shake.
Take note of the phenomenon.

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2.3.4. Starch is produced during photosynthesis
- Use black paper to partially shade the leaves (on
the tree) for 2 days (Figure 7). Break the leaves,
transfer the leaves with tongs into a test tube
containing 70 alcohol in a beaker of boiling water,
and heat until the leaves lose their green color.
- Wash the leaves with water and spread the leaves
on a petri dish. Pour the Lugol’s solution onto the
leaves and shake to stain the leaves. Use tissue paper
to absorb excess Lugol’s solution.
- Take note of the phenomenon.
Figure 7. Starch test

3. REPORT
1. Identify the composition of the pigment mixture on the chromatographic paper. Define Rf. Why
can these pigments separate and move to different locations on the chromatographic paper? Include
a signed copy of the report.
2. Describe, draw (take photos), and explain all phenomena in experiments 2.3.2, 2.3.3, 2.3.4.

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