Biology I

Laboratory
- Manual

Biology Department
2022
First Semester Course outline 2022-2023
Week 1 28- 30 /8 Orientation week
Week 2 4 8 /9 Introduction to lab apparatus and lab safety
Week 3 11-15 /9 Introduction to microscope
Week 4 18-22 /9 Chemical composition of living organisms
Week 5 25-29 /9 Preparation of check cell slide
Week 6 2- 6/ 10 Preparation of onion cell slide
Week 7 9-13 / 10 Histological Examination of Tissues 1
Week 8 16-20/ 10 Histological Examination of Tissues 2
Week 9 23-27/ 10 Quiz 1
Week 10 30/10 - 3 /11 Cell division (Mitosis & Meiosis)
Week 11 6- 10/ 11 Blood Types
Week 12 13 -17 /11 DNA extraction
Week 13 20 -24 / 11 Enzymes
Week 14 27/11- 1/12 Mid term vacation
Week 15 4 8 /12 Quiz 2
 Introduction to
lab safety, first aid & lab apparatus
 Laboratory Safety

Read instructions carefully before starting the lab. Be sure to ask

your teacher about anything that you do not understand.

Keep your lab area clear of any materials not needed for the
experiment.

Never eat or drink in the lab.

Broken glass and sharp objects do not go in trash cans.

 Laboratory Safety
Never point the end of a test tube being heated at
yourself or others.

Be sure your hands and your lab area are dry before
using electrical equipment.

Unplug cords by pulling the plug and not the cord.

Unplug all electrical equipment at the end of the lab

period.
 Laboratory Safety

Keep all long hair tied back.

Wear your lab coat.

Foot wear that completely covers the foot is required.

 In case of an Emergency

Know the locations of:

fire extinguisher, fire blanket, body shower, eyewash station and first aid kit.
 First Aid

Injury: Burns
What to do:

Immediately flush with cold water until burning sensation is lessened.

 First Aid

Injury: Cuts, bruises

What to do:
Do not touch an open wound without safety gloves.
Pressing directly on minor cuts will stop bleeding in a few minutes.
Apply cold compress to bruises to reduce swelling.
 First Aid

Injury: Fainting
What to do:
Provide fresh air.
Have the person recline so that his head is lower than his body.
 First Aid

Injury: Eyes
What to do:
Flush eyes immediately with plenty of water for several minutes.
If a foreign object is lodged in the eye, do not allow the eye to be rubbed.
 First Aid

Injury: Spills on the skin

What to do:
Flush with large quantities of water.
For acid spills, apply baking soda solution.
For base spills, apply vinegar or boric acid.
 First Aid

Injury: Electrical shock

What to do:
Shut off the current at the source.
Remove wire with rubber gloves.
Alert the teacher immediately.
 Laboratory Apparatus
There are many apparatus that are used in a laboratory:

Microscope:
Is an optical instrument that uses a lens or a combination of lenses to produce
magnified images of small objects, especially of objects too small to be seen
by the unaided eye.
 Laboratory Apparatus

Microscope slide:
The microscope slide is a flat piece of glass, used to serve as a
platform for viewing objects under the microscope.

Cover slip:
Is thin flat piece of transparent material, usually square or
rectangular, which is placed over objects held by the slide.
 Laboratory Apparatus

Dissecting set:
a set contains tools to perform dissections.

Spatula:
used for moving small amounts of powder from place to place.

Scalpel:
It is a small, sharp knife used in performing dissections.

Forceps:
Are used for grasping and holding objects.
Introduction to Microscope
 Introduction to Microscope

Light microscopes:
a biology laboratory instrument, that uses visible light to detect
and magnify very small objects and enlarge them.

There are two types of light microscope:

1. Simple light microscopes
Has one lens of magnification

1. Compound light microscopes

Has two lenses of magnification
 Parameters in Microscopy

Two parameters are especially

important in microscopy:
1. Magnification: The ability of a
microscope to produce an image of an
object at a scale larger (or even smaller)
than its actual size.

2. Resolution: The ability of a microscope

to distinguish detail.
 Compound Light Microscope
The compound light microscope is used to observe small structures such as
cells and tissues.

The term compound refers to the two types of lenses (ocular and objective)
that are used to magnify the image.

The compound microscope can magnify the images up to

approximately 3,000 times, depending on the magnifying power
of the lenses.
 Care of a Compound Microscope

The compound microscope is an expensive instrument that must be handled

appropriately:

Pick up the microscope with two hands, one holding the arm and the other
supporting the base.

Carry the microscope upright so that a lens or eyepiece

does not fall out, and carefully place the microscope on
the lab table in front of you.
 Parts of the Microscope

Microscopic parts Functions

Base Supports the microscope.

Arm Connects the base to the head.

Head (or body tube) Contains the ocular lenses and the rotating nosepiece with the objective
lenses.
Ocular lenses Observe the microscope slide.
pointer Identify a specific area on the slide.
Magnify an object, 4X (Scanning), 10 X (low- power), 40 X (high-
Objective lenses power) and 100 X (oil immersion).

Stage The microscopic slide is placed

 Parts of the Microscope
Microscopic parts Functions

Stage clips Hold the slides in proper place.

Adjuster knobs Moves the slide side to side and the other forward and backward.

Coarse focusing and moves the stage up and down.

Coarse focus knobs

Fine focus knob Precision focusing.

Condenser Condenses the light through the specimen on the slide above.

Iris diaphragm Regulates the amount of light that passes through the condenser.

Light source is usually built into the base of the microscope

 Calculate Magnifications

Total magnification is determined by multiplying the ocular lens power

and objective lens power.

Objective lens Ocular lens Total magnification (X)

4X 10 X 40X
10X 10 X 100X
40X 10 X 400X
 Objective lenses
Arm
Ocular
lens
Stage clips
Head

Coarse focus knobs

Stage

Condenser Fine focus knob

Iris diaphragm

Adjuster knobs
Light source

Base
Chemical composition of living
organisms
 Level of Organization

Living organism have a high degree of organization.

Organization levels of living organisms are:

1. Atoms 5. Tissues
2. Molecules 6.Organs
3. Organelles 7. Organ systems
4. Cells 8. Organism
Level of Organization
 Organic compounds

There are four classes of macromolecules (or organic compounds)

within living cells:

1. Carbohydrates
2. Proteins
3. Lipids
4. Nucleic acids
 Detection of Carbohydrates, Proteins & Lipids

Objectives: Materials:
Test tube
Starch
After completing this exercise you Water
will be able to: Water bath
Diluted iodine solution
Identify the major classes of organic Potato juice
molecules. Microscope
Providing examples of each Grape juice

Egg albumin
Sodium hydroxide (20%).
Copper sulfate (1%).
Filter paper
Oil
 Detection of Carbohydrates
Carbohydrates are major source of energy for cells.

Three classes of carbohydrates:

1- Monosaccharides
2- Disaccharides
3- Polysaccharides

In this lab we will detect starch grains (polysaccharide).

 Detection of Carbohydrates (polysaccharide)

There are two methods to detect starch:

 1- Detection of starch by microscopic examination
Procedure:
1. Put a drop of potato juice in the center
of a clean slide.

2. Add two drops of water by a dropper then

place a cover slip.

3. Examine under microscope using low (4x)

then high (40x) powers.
1- Detection of starch by microscopic examination

Observation:
 2- Detection of starch by chemical methods
Procedure:

1. Put a little amount of starch in a test tube.

2. Add 5 ml of water.

3. Heat up in a water bath for a few minutes.

4. Shake well until all starch dissolves in the water.

5. Add two drops of diluted iodine solution and shake.

 2- Detection of starch by chemical methods

Observation:
 Detection of Carbohydrates (Monosaccharides)

Procedure:

1. Put 1 ml of grape juice in a test tube using a dropper.

(grape juice rich in glucose, Which is a monosaccharide)

2. Add 5

3. Heat up in a water bath until boiling.

4. Take the tube out and put it in a tube rack

for few minuets.
 Detection of Carbohydrates (Monosaccharides)

Observation:
 Detection of Proteins by Biuret Test
Proteins are complex molecules that play many critical
roles in the body.
Procedure:
1. Put few drops of egg albumin in a test tube.

2. Add 1 ml of sodium hydroxide (20%).

3. Add a drop of copper sulfate (1%).

4. Take the tube out and put it in a tube rack

for few minuets.
 Detection of Proteins by Biuret Test

Observation:
 Detection of Lipids by a Filter Paper
Lipids are hydrophobic compounds.

Procedure:

1. Take a filter paper and add one drop of water on one end and a drop of oil or lipid on the
other end.
2. Expose to light and notice.
 Detection of Lipids by a Filter Paper

Observation:

transparent permanent stain

 Observation

Macro- Example color

INDICATOR Molecules
Iodine
Solution

Solution

Copper
Sulfate
Solution
 Cell Staining
Preparation of Cheek Cell Slide
Cell Staining & Preparation of Cheek Cell Slide

Objectives Materials
After completing this Microscope
exercise you will be able to: Toothpick
Define cell staining Cover slip
Identify the cell parts Water
Observed in cheek cell slide Methylene Blue
Write the function of each Microscope Slides
part of the cell Plastic pipette
Tissue paper
 Cell Staining

Many samples, particularly cells, can appear quite transparent under the
microscope. The internal parts of the cell, the organelles, are so
transparent that they are often difficult to see.

Biologists have developed a number of stains as methylene blue, iodine,

hematoxylin and eosin.

Why do we have to stain cells?

To enhance visualization of the cell or certain cellular components under a
microscope.
 Preparation of Cheek Cell slide
Observing human cheek cells under a light microscope is a simple way to
quickly view a human cell structure.

Cheek cells are epithelial cells that line the interior surface of our mouth.

Most of the organs inside the body are lined with a layer of epithelial cells.

Scientists choose specific stains when they want to look at a particular part
of a cell so before you start the experiment, decide which stain you will use.
 Slide Preparation Procedure
1. Gently scrap the inside of your cheek by using the
flat edge of the tooth pick.

2. Place a drop of water on a microscope slide.

3. Gently swirl the end of the toothpick into the drop of

water.
 Slide Preparation Procedure

4. Place a small drop of methylene blue dye onto the

drop of water and cheek cells.

5. Place a cover slip over the slide by placing the

bottom edge in the water drop and holding
it at a 45 degree angle to the slide, If air bubbles
are seen gently press on the cover slip to release
the bubbles.
 Slide Preparation Procedure

6. Pull excess dye and water out from underneath the cover
slip by allowing a paper towel to touch one side of the
cover slip.

7. Set the magnification of your microscope to 40X.

Look at the stained slide under the microscope.
 Observation

Observe the major components of the human epithelial cheek cell.

The irregular shape of cell, cell membrane, nucleus and cytoplasm.

Preparation of Onion Cell Slide
 Plant Cell

Plant cells are eukaryotic cells.

Plant cell structure includes various components known as cell organelles
that perform different functions to sustain itself.
Some of the organelles present in plant cells are different from other
eukaryotic cells, for example:
1. Plastids
2. Vacuoles
3. Cilia
4. Cell Wall
 Preparation of Onion Cell Slide

Objectives Materials
Microscope
After completing this Cover slip
exercise you will be able to:
Onion
Identify the cell parts. Iodine Tincture
Observed in onion cell slide. Kitchen Knife
Write the function of each Plastic pipette
part of the cell. Cutting Board
Forceps
 Slide Preparation Procedure
Squamous cells of onion epidermis show many feature
found in non green plant cells.
1. Cut a piece of one of the base of fatty onion leaves.

2. Remove away a small thin piece of epidermis with forceps.

3. Put this piece of epidermis on a drop of diluted iodine

solution on a clear slide (try to spread the onion epidermis).
 Slide Preparation Procedure

4. Cover the sample by clean glass cover.

5. Place the microscope slide onto the stage and

secure with the stage clips.

6. Observe the onion cell under microscope.

 Observation

Microscopic examination of squamous epidermis of onion (plant cell),

allow as to identify:

1. Thick cell wall surround the cell.

2. Large vacuole.
3. Nucleus inside the cell.
Histological Examination of Tissues
 Types of tissues in human body

Types of tissues in human body

Epithelial tissue

Connective tissue

Muscular tissue

Nervous tissue
 Epithelial tissues

Features:
They appear as sheets.
Consist of single or multiple layers.
They have different shape.
They always rest on a basement membrane.
Covers surfaces, lines cavities, and forms glands.

Functions:
Absorption
Secretion
Protection
 Epithelial Tissues Shape & Arrangement

Simple Stratified

A - Squamous
Shapes of epithelial tissues

epithelium

B- Cuboidal
epithelium

C- Columnar
epithelium

D- Transitional
epithelium
 Simple Cuboidal Epithelium

Structure:
It consists of a single layer of cube-like (cubical) cells lying on a basement
membrane.
The nucleus is usually rounded and located in the center of the cell.
 Examine a slide of Simple Cuboidal Epithelium

Location: lines the lumen of the kidney tubules.

Function: Absorption
Examine a slide of Stratified Cuboidal Epithelium

Structure:
This epithelium is composed of multiple layers of cuboidal cells.

Location: Lining of salivary gland ducts.

Function: Secretion.
 Connective Tissues
Connective tissue is the most abundant primary tissue in
the body and has a variety of functions.

and other tissues together.

Connective tissue is made up of two main components:

Cells and extracellular matrix.

Extracellular Matrix is composed of ground substance

and fibers.
 Types of Connective Tissues

Areolar tissue
Types of Connective tissues

Loose Connective tissue

Dense Connective tissue Adipose tissue

Cartilage

Bone

Blood
 Examine a slide of Areolar Connective Tissue
Structure:
It is the most widely distributed of all connective tissue types.
It consists of a meshwork of fibers with many connective
tissue cells in between the fibers.

Location:
Around blood vessels.

Function:
To help keeping the vessel in place.
 Examine a slide of Adipose Connective Tissue
Structure:
Fat cells called adipocytes. Each adipocyte occupies most of the volume of
the cell, its cytoplasm, nucleus and other components are pushed towards the
edges of the cell.

Location:
Under skin, around kidneys and heart.

Function:
Protect internal organs, stores energy.
 Examine a slide of Dense Connective tissue
(fibrous tissue)
Structure:
Is composed mainly of large amount of collagen fibers arranged in bundles.
the collagen fibers are arranged in parallel rows with the fibroblast located
between the fibers.
 Examine a slide of Dense Connective tissue
(fibrous tissue)
Location:
Found in tendons (connect muscles to bone)

Function:
Support, strength and mobility of muscles and bone.
 Examine a slide of Cartilage
Cartilage cells are called Chondrocytes. These are large,
roughly spherical cells, each containing a big centrally placed
nucleus. The chondrocytes are located within small cavities in
the matrix. These cavities are called lacunae.
Location: Trachea.
Function: Support and keep constant opening of its lumen and
prevent its collapse.
 Examine a slide of Bone
Structure of bone:
Haversian system consisting of:
1. Haversian Canal: a central canal.

2. Lamellae: layers or rings of a strong

matrix.

3. Lacunae: small spaces between the

lamellae surrounding the Haversian canal
which contain the bone cells (Osteocytes).
 Examine a slide of Bone

4. Canaliculi: fine canals connecting lacunae together, provide routes by

which nutrients can reach the osteocytes.
 Examine a slide of Bone

Function: Bones are a storage area for calcium and provide support and
protection for the body.
 Examine a slide of Blood

The blood composed of plasma and blood cells.

Types of blood cells:

1. Erythrocytes (RBCs)
2. Leukocytes (WBCs)
3. Platelets.
 Examine a slide of Blood

Function: Transport nutrients and wastes. Red blood cells carry oxygen.
 Cell Division
( Mitosis & Meiosis)
 Cell Division ( Mitosis & Meiosis)

Cell divison occur in all living organisms.

There are two main type of cell divison in eukaryotic cells:

1. Mitosis
2. Meiosis
 Mitosis (Mitotic cell division)

Is a process of cell division which results in the production of two daughter

cells from a single parent cell.

The daughter cells are identical to each other and to the original parent cell.

Mitosis maintains a constant amount of genetic material from one generation

to another.
Stages of Mitosis
 Significance of Mitosis

1. Growth: The number of cells within an organism increases

by mitosis and this is the basis of growth in multicellular organisms.

2. Cell Replacement: Cells are constantly dying and being replaced by

new ones. When damaged tissues are repaired, the new cells must
be exact copies of the cells being replaced so as to retain normal
function of cells.
 Stages of Mitosis

Interphase Prophase Metaphase Anaphase Telophase

Examine the prepared slide of root tip of onion plant
 Meiosis

Meiosis is a type of cell division that reduces the number of chromosomes

in the parent cell by half and produces four gamete cells.

Significance of Meiosis:
It is responsible for formation of sexual cells or gametes (egg and sperm
cells) that are responsible for sexual reproduction and also produces
genetic variation by way of the process of recombination.
Stages of Meiosis
Examine the prepared slide of animal cell
Blood Types
 Blood Types
Human Blood is about 45% cells and 55% plasma

Blood cells
including red blood cells, white blood cells and
platelets

Plasma
including water, gases, nutrients, wastes, ions,
hormones, enzymes, antibodies and other proteins.
 Blood Types
On the surfaces of your red blood cells are one or more antigens that will
cause their agglutination if exposed to the complementary antibodies.

Agglutination is the clumping of erythrocytes. This could theoretically occur

during blood transfusion if there is transfusion of incompatible blood.

That causes the destruction of donor erythrocytes and perhaps the death of the
patient when clumping cells block blood vessels.
 Blood Types

For these reasons, blood used for transfusion is very carefully matched for

The Blood type you belong to depends on what you inherited from your
parents.
Agglutination
 ABO Blood Typing System

There are four kinds of blood types:

A, B, AB or O
Blood Group A
If you belong to the blood group A, you have A antigens on
the surface of your red blood cells and B antibodies in your
blood plasma.

Blood Group B
If you belong to blood group B, you have B antigens on the
surface area of your red blood cells and A antibodies in your
blood plasma.
 ABO Blood Typing System

Blood Group AB
If you belong to blood group AB, you have A and B antigens
on the surface of your red blood cells and no AB antibodies at
all in your blood plasma.

Blood Group O
If you belong to blood group O, you do not have either A nor
B antigens on the surface area of your red blood cells but you
have both A and B antibodies in your blood plasma.
ABO Blood Typing System
 Rhesus factor (Rh)

Rh grouping system is a system for classifying blood groups according to the

presence or absence of the Rh antigen.

Those who have it are called Rh+.

Rh-.
 Blood Group Test

Materials:
Objectives:
Sterilized lancet
Find out your
Clean glass slide
own blood group
Alcohol swab
Recognize Rh factor
Antiserum A , B, and D.
 Procedure

1. Clean a finger.

2. Make a puncture on a fingertip with a sterile lancet

3. Place three small drops of blood on the slide.

 Procedure

4. Mix the blood with three different antiserum,

these antisera contains different antibodies,
A, B & D antibody.

5. Then you observe reaction that is happening, in

which mixture has agglutination occurred?
 Example

Anti Sera A Anti Sera B Anti Sera D

 Observation
Antiserum Blood Blood Blood Blood
sample sample sample sample
A+ B- AB+ O-

D
Catalase Enzyme
 Catalase Enzyme

Enzymes are biological catalysts that increase the rate of chemical reactions
without being used up.

They are large protein molecules that are folded into complex shapes that
allow smaller molecules called substrate to fit into them.

The place where these substrate molecules fit is called the active site.
How Enzyme Work
 Catalase Enzyme
All living tissues contain catalase enzyme.
They found in food as potato and liver.
It is one of the fastest enzymes in the cell that used for removing Hydrogen
Peroxide H2O2 from the cell.
Hydrogen Peroxide is produced as product of many normal cellular
respiration. If the cells did not break down the Hydrogen Peroxide, they would
be poisoned and die.
H2O2 can be converted to oxygen and water by catalase enzyme.
 Catalase Enzyme

Objectives Materials
Raw chicken liver
After completing this exercise Onion
you will be able to: Potato
Hydrogen peroxide
Identify the main function of (3%H2O2 )
catalase enzyme. Forceps
Scissors
Test tubes
 Procedure

Place a small piece of liver, potato and onion separately in test tubes and
label them.

Add 5 ml of 3% H2O2 in each test tube.

Observe the reaction.

 Observation

Tissue observation
Onion
Potato
Liver
DNA Extraction
 Deoxyribonucleic acid (DNA)
Deoxyribonucleic acid, more commonly known as DNA.

DNA is located in the cell nucleus that carries the genetic information
 DNA Structure
DNA is a double helix.

DNA is composed of two polynucleotide chains

joined by hydrogen bonding between bases,
twisted into a helical shape.

The sugar-phosphate backbone is on the outside.

The nitrogenous bases are perpendicular to the

backbone in the interior.
 Deoxyribonucleic acid (DNA)

The building unit of DNA is nucleotide .

Components of a nucleotide:
Made of three parts:
1. Pentose sugar (deoxyribose)
2. Phosphate group
3. Nitrogen bases
 DNA Extraction
DNA Extraction is the removal of deoxyribonucleic acid (DNA) from the
cells.

Basic steps of DNA Extraction:

1. Break open (lyses) the cells.

2. Degradation of cellular proteins.

3. DNA is the precipitated by mixing with cold ethanol or isopropanol.

 DNA Extraction

Objectives Material
Strawberries
Understand the process of Filter paper
DNA extraction. Extraction buffer
Small funnel
90% Ethanol ice-cold
20 ml Graduated cylinders
10ml test tubes
Zip-lock bags
 Procedure

1. Place two strawberries in a zip-lock bag,

press the air out, and then seal it.

2. Softly mash the strawberry with your fingers

for 1-2 minutes until it becomes a juice puree
careful not to break the bag!!
 Procedure

3. Add 10mL of the provided extraction buffer and then press

the air out of the bag and seal.

4. Mash the strawberry carefully for 1 minute without creating

many bubbles.
 Procedure
5. Place test-tube in a cup. Put the funnel on
top of the test-tube. Place the filter paper on
top of the funnel.

6. Open the bag and drain carefully the strawberries

on top of the filter paper to fill the test-tube with
juices. The juice will drain through the filter paper
but the chucks of strawberries will not pass through
into the test-tube.
 Procedure

7. Tilt the test-tube and pour in an equal amount of

alcohol, through the funnel and down the sides of
the test-tube. This will allow for better separation of
the DNA.

8. Dip the loop or glass rod into the tube where

the strawberry extract and ethanol layers come
into contact with each other.
 Observation

9. Gently remove the stirring rod and examine what the DNA looks like.