Unit 4

CELL BIOLOGY
Cell: is the smallest structural and functional units of life.
Cell Theory: is the scientific theory which describes the nature, structure, function,
origin and properties of cells.
Ideas stated under cell theory:
✓ Cells are the smallest structural and functional unit of life.
✓ All living Organisms are made up of one or more cells.
✓ All cells come from pre-existing cells by cell division. That means, “cells do
not arise spontaneously from non- living matter”.
How did the modern cell theory develop?
✓ The discovery of cells had related to the development of microscope because
no one observed them until microscope were invented.
✓ A number of scientists contributed their own work in developing ideas about
cells under the cell theory. Some of these scientists are:
1. Robert Hooke (1665)
✓ He saw tiny structures in the cork plants and he called them “Cellular” which
means “smaller inner room” using his primitive compound microscope with a
magnification power of 30X.
✓ He saw smaller hallow inner rooms in the cork plant and he called them cells.
✓ He was the first person who used the term cell to describe the smaller inner
units that he observed in the cork plant.
2. Anton Van Leeuwenhoek (1674)
✓ He observed small living cells (unicellular moving organisms) which capable
of moving in a drop of water. They were Protoctista and today what we called
them protozoa.
✓ He discovered small living single celled organisms and he called them
“animalcules” which means small moving organisms.
✓ He also observed “bacteria” from his teeth and he called them “tiny
animalcules “
3. Rene Dutrochet (1824)
✓ He was the first man who stated the concept of cell theory.
✓ He stated the followings ideas about cells in his cell theory:
• All organisms are made up of cells.
• The growth of all organisms is due to the increase in volume of cells by
mitosis.

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4. Matthias Schneider and Theodor Schwann (1839)
✓ They forward the first clear cell theory and stated the following ideas in their
cell theory:
• Cell is the smallest unit of structure and physiology in living things.
• Cell retains a dual existence as:
❖ a distinct entity which is capable of independent existence
❖ a “building block” in the formation of organisms
• Cells are formed spontaneously by free-cell formation (spontaneous
generation). This idea of cell has now been proved wrong.
5. Rudolf Virchow (1858)
✓ He declared that: Omnis cellula e cellula which means that one cell can arise
from another cell like it by cell division.
✓ He completed the first accepted version of the cell theory by adding a new idea
about cell. Like the idea that “All cells come from pre-existing cells”.
Some Important Ideas stated under the Modern cell theory:
✓ Cells are the smallest structural and functional unit of life.
✓ All living things are made up of one or more cells.
✓ All cells come from pre-existing cells by cell division. That means “cells do
not arise spontaneously from non- living matter”.
✓ All cells have basically the same chemical composition.
✓ They have DNA (genetic materials) which passed from cell to cell during cell
division.
✓ All cells contain hereditary materials or information which used to make
instructions to different body traits and growth.
✓ All the energy flow occurs within cells. That is, all kinds of metabolisms occur
within cells.
Types of cells
✓ Biologists classified cells in two main groups; based on their nature, appearance,
structure, metabolism and reproduction. These are:
A) Prokaryotic cells B) Eukaryotic cells
1) Prokaryotic cells
✓ These are the first types of cells formed when life was first evolved on earth.
✓ They named from Greek words meaning “before nucleus”.
✓ They don’t have membrane bounded nucleus. That is, they do not have “true
nucleus”.

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 ✓ But they have a nuclear region called “nucleoid” which contains a
genetic material (DNA) however; it is not enclosed with nuclear
membranes.
✓ They do not have organelles (internal membrane systems). They have
only the small “70’s ribosome’s”.
✓ They are the simplest and primitive form of cells because they do not
have:
➢ Organelles (or membrane bounded structures).
➢ Much more division of labor. Because they no specialized regions
(Organelles) to perform different metabolisms of the cells in
separate places or regions.
➢ An organized genetic material in the form of chromosome. That
is, they have only a single circular DNA as genetic material.
✓ All kinds of organisms in the kingdom monera are prokaryotic cells
Bacteria - They are Prokaryotic cells (or organisms)
Blue green algae - They are grouped in kingdom monera.
(Cynobacteria)
2. Eukaryotic cells
✓ The word eukaryotic comes from the Greek word meaning true nucleus.
✓ Eukaryotic cells have membrane bounded nucleus (or true nucleus) which
contains the genetic material in it in the form of chromosomes.
✓ They have also many membranes bounded organelles each has its own
specific function.
✓ They have organelles in their cytoplasm such as nucleus, mitochondria, Golgi
apparatus and plastids (chloroplasts) in photosynthetic Eukaryotes
✓ They are much more complex and advanced cells than prokaryotic cells because
eukaryotic cells have: -
• Different organelles and each perform specific function (metabolism).
• Much more division of labor due to the presence of various organelles.
• Organized genetic material (DNA) in the form of chromosomes
✓ All cells of fungi, protozoa, algae and multicellular organisms such as animals
and plants are eukaryotic cells.
✓ All cells in kingdom Plantae, Animalia, fungi and Protista are Eukaryotic cells.

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The difference between Eukaryotic and Prokaryotic cells
Feature Prokaryotic Cells Eukaryotic Cells
Size 1-10µm 10-100µm
-It has no membrane bounded -It has Nucleus surrounded by
Nucleus Nucleus nuclear membrane
-It has no definite Nucleus -It has definite Nucleus
-It has a continuous loop like -It has a linear form of DNA
DNA -It is surrounded with Histone
DNA
-It is not surrounded with protein proteins to form chromosome
to form chromosomes
Mitochondria Absent Present
-Absent, but some of them can -Present, in plant cells and Algal
Chloroplasts make photosynthesis with cells only
Chlorophyll
It has 70S Ribosomes which are -It has 80S Ribosomes which
Ribosomes
smaller than in Eukaryotic cells are larger than Prokaryotic cells.
Always present, often made from -Present in Plant cells, Algal
Peptidoglycan not Cellulose cells and fungal cells
-Cellulose cell wall in plant
Cell Wall
cells
-Cell wall made from Chitin in
Fungal cells

The Origin of Eukaryotic cells

✓ There is Endosymbiont theory that explains the origin of Eukaryotic cells.
✓ It is the theory which states that “the modern Eukaryotic cells” were formed
when several primitive prokaryotic cells “got together”.
✓ the evolutionary line of Eukaryotic cells is explained as follows:-
• The plasma membrane of the ancestral prokaryotic cells became more and
more “in folded and became an extension of internal membrane system.
• This would eventually evolve into the endoplasmic reticulum (EPR) of
Eukaryotic cells. These membranous prokaryotic cells engulfed other
smaller prokaryotic cells.
• These prokaryotic cells engulfed either photosynthetic prokaryotic cells or
prokaryotic cells that had a better respiring to release energy. As a result of
this, two evolutionary lines were formed this time.

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 1. If the engulfed prokaryotic cells had a better way of respiring to release energy,
then they would evolve into the mitochondria of eukaryotic cells.
✓ These cells that contained the mitochondria were heterotrophic Eukaryotes and
they were the ancestors (forerunners) of:
❖ Animal cells
❖ Fungal cells
❖ Protoctistants (protozoa cells)
2. On the other hand, if the prokaryotic cells with primitive mitochondria engulfed
other small prokaryotic cells that could photosynthesis, then they were evolved
into chloroplasts.
✓ These cells that contained chloroplasts were autotrophic Eukaryotes and they
were the ancestors of plant cells and algal cells.
The main groups of organisms (living things)
The Group of organisms which consists of Prokaryotic cells are:
A) Archaebacteria:
✓ Are the oldest organisms on earth which consists of unicellular prokaryotic
cells. They evolved when conditions on earth were very harsh.
✓ They are still found in a very hot environment and large concentration of
chemicals like Sulphur dioxide and methane.
B) Eubacteria: are true or advanced bacteria consist of unicellular prokaryotic cells.
The group of organisms which consists of eukaryotic cells are:
1. Protists (kingdom protista): consists of either unicellular or multicellular
Eukaryotic cells.
2. Fungi (kingdom fungi): consists of multicellular Eukaryotic cells.
3. Animals (kingdom Animalia): consists of multicellular Eukaryotic cells.
4. Plants (kingdom plantae): consists of multicellular Eukaryotic cells.
Parts of the cell and their Functions
Some of the main parts of cells are cell membrane, Nucleus and Organelles such as
mitochondria, ribosomes, lysosomes, chloroplasts and Golgi – complex.
1) Cell membrane/plasma membrane
✓ It is the membrane that isolates the cell from its environment.
✓ Is the membrane that surrounds and encloses all the component of the cells.
✓ It separates the cell’s internal contents from extracellular environment.
✓ It helps specialized metabolism processes such as photosynthesis and aerobic
respiration to be taken place in a separate living cell.

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 ✓ It is commonly called plasma membrane. It contains molecules that allow the
cell to be recognized by:
➢ Hormones, immune system and drugs (in animals).
➢ Growth regulator substances, such as axons (in plants)
✓ It acts as receptor sites or chemical signals to recognize hormones and
neurotransmitter. It is used as cell signal.
✓ It controls the exchange of substances between the cell contents and
environment.
✓ Plasma membrane allows substances to move in and out of the cell by:
❖ Passive processes ❖ Active processes
➢ Simple diffusion ➢ Active transport
➢ Facilitated diffusion ➢ Endocytosis
➢ Osmosis ➢ Exocytosis
The Structure of Cell membrane (plasma membrane)
The basis or skeleton of cell membrane is phospholipid bilayers.
✓ Cell membranes are mainly made up of lipids and proteins
✓ Phospholipids bilayer is the main fabric (knitted materials) of the plasma
membrane.
There are two molecular models of the plasma membrane to show the
arrangements of phospholipids and proteins in it. These two models of
membranes are: -
A) The Davison-Danielli Model
✓ Davison and Danielli knew that both proteins and phospholipids
were involved in the structure of plasma membranes.
✓ They suggested a kind of ‘sandwich’ model of protein and
phospholipids.
✓ This model is also known as a sandwich model of phospholipids and
proteins.
✓ This model of cell membrane proposed that the phospholipids bilayer
is sandwich between two layers of proteins.
✓ It is unstable model of membrane because the phospholipids heads
separated from the water by such types of protein arrangement.
✓ This model could not adequately explain all the new evidence. This
model therefore had to be rejected.

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 B) The fluid mosaic model
✓ S.Singer and G.Nicolson proposed the revised model of plasma membrane
known as the fluid Mosaic Model.
✓ They retained the idea of phospholipid bilayer, but they rejected the
sandwich arrangement of the Davison- Danielli model. Instead, they
suggested that proteins were ‘studded’ into the bilayer at different points.
✓ They proposed that protein’s membrane is dispersed and individually
inserted into the phospholipid bilayers.
✓ They suggested that proteins were “studded “into the phospholipid bilayers
at different points.
✓ This model stated that the hydrophilic portions of protein membrane
protruding far enough from the bilayers to be exposed to water. This
maximizes the contact of the cell to hydrophilic region of protein and
phospholipids with water.
✓ This model stated that cell membrane is a mosaic of protein molecules
immersed in fluidic bilayers of phospholipids and hence it is called “The
Fluid Mosaic Model “.
✓ This model suggested that the arrangement was not static, but was fluidic
and constantly changing.
✓ This model of plasma membrane (molecular arrangement of protein and
phospholipid) is more stable form of model than the Davison- Danielli
model.
Why we call this model is the fluid mosaic model?
A. It is fluidic because:
• The phospholipids in membrane can move and change its position.
• There is a lateral and flip flop movement of phospholipids in the plasma
membrane.
• The membrane has fluidity nature due to the unsaturated fatty acids (lipids) of
membrane.

B. It is mosaic because the arrangement of proteins in the membrane gives a

patchwork appearance (or the proteins looks mosaic).
The key features and molecules in the plasma membrane of the fluid mosaic model
of the cells:
→ The phospholipids bilayers are the basis for the plasma membrane.

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→ Integral proteins (intrinsic proteins): are proteins that span both phospholipid
layers in a plasma membrane. They also known as trans-membrane proteins.
→ Integral proteins play an important role in moving substances across the
plasma membrane. There are three main types of these transport proteins.
These are:
A. Channel proteins:
✓ Are integral proteins with channels (ions pores) that allow specific ions to
pass through the plasma membrane.
✓ There different channel proteins to allow different ions.
B. Carrier proteins:
✓ Are integral proteins that move large molecules (medium sized particles) through
the membrane by facilitated diffusion or active transport.
✓ The carrier proteins that involved in the active transport are often referred to as
pumps.
C. Peripheral proteins (extrinsic proteins): are proteins that span only one of the
two phospholipid layers. They have a range of functions:
• some are used as enzymes and some others anchor integral proteins to the
cytoskeleton.
❖ Glycolipids :–
✓ Are lipid molecules that have carbohydrate chains attached on them.
✓ The carbohydrate component of each Glycolipids can be cell specific and so
they allow the identification of the cell by the immune system.
✓ They serve as cell signals to other cells.
✓ Are also serve as receptor sites for chemical signals like hormones,
enzymes, immune system and drugs.
❖ Glycoproteins:
✓ are proteins that have carbohydrate chains attached on them.
✓ The carbohydrate component of each glycoprotein can be cell specific
and so they allow the identification of the cell by the immune system.
✓ They serve as cell signals to other cells. They also serve as receptor sites
for chemical signals.
❖ Cholesterol:
✓ Is the type of lipid that is slightly polar at one end and so it helps polar
molecules through plasma membrane.
✓ It increases the flexibility and stability of the membrane.
✓ It reduces the fluidity of the membrane.

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How do substances move through the plasma membrane of cells?
There are two processes by which substances cross plasma membranes. These are:
1. passive processes:
• Depend on the kinetic energy of the particles of the substances.
• They also depend on the concentration gradients of the particles.
• They do not need extra energy (ATP) from the cell’ respiration
(metabolisms).
Types of passive processes:
A) Simple diffusion: Is the movement of particles from a high concentration area to a
lower concentration area.
→ This diffusion particles occurs when there is a concentration difference between
the two sides of the plasma membrane.
→ The particles in the fluids ( liquids and gases ) are free to move and so they have
mostly high kinetic energy to drives diffusion.
→ The particles that move in and out of the cell through plasma membrane by simple
diffusion are: - Small particles
-Lipid soluble (non- polar fat-soluble molecules)
- Non –charged molecules
→ The following particles cannot move across plasma membrane by simple diffusion.
• Ions - charged particles
• Sugars and amino acids - they are not lipid soluble and they are large
particles.
• Proteins - large particles
The rate of diffusion that occurs through the plasma membrane is influenced by the
following factors:
❖ The concentration gradient
❖ The diffusion distance ---- the thickness the membrane
❖ The surface area of the membrane.
 All these factors are related in the equation called Fick’s law of diffusion. It
explains how these features influences the rate of diffusions.
Rate of diffusion α surface area of membrane x concentration difference
Diffusion distance
✓ The rate of diffusion is also influenced by temperature.
✓ Diffusion occurs in a faster rate at a higher temperature because the particles
have more kinetic energy.

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C) Facilitated diffusion: Is the movement of particles from a higher concentration area
to a lower concentration area with the help of channel proteins (ion pores) or carrier
proteins.
❖ The carrier proteins undergo a conformational change to move larger particles
through the cell membranes.
→ Like simple diffusion, the particles move from high concentration gradient to low
concentration gradient with the help of protein transport (carrier proteins &
channel proteins).
→ The rate of facilitated diffusion is affected by the same factors to the simple
diffusion, except the factor of surface area of the membrane.
• Concentration difference- Diffusion distance
• Number of channel proteins or carrier proteins
D) Osmosis:
→ Is the diffusion of water by which water moves across a partially permeable
membrane.
→ Is the movement of water which depends on water potential.
→ Is the movement of water from a system which has high water potential (less
negative Ѱ) to a system which have low water potential (more negative Ѱ) across
plasma membrane. The symbol for water potential is the Greek letter Ѱ (psi).
➢ Water potential (Ѱ) is the measurement free water molecule in the system.
➢ Ѱ(water potential) is measured in units of pressure-Pascal(pa)
➢ Pure liquid water has a higher water potential than any other system. It is
defined as zero.
 Ѱ (pure water) = 0 Pa
✓ All the other systems (such as cytoplasm of cells and any solutions) have a
lower water potential than water.
✓ The water potential of cells and any solutions have negative value.
✓ Cytoplasm of Cells - have more negative water potential.
✓ Any solutions - lower water potential
✓ The rate of osmosis is influenced by the same factors as simple diffusion.
✓ Surface area of the membrane
✓ Difference in water potential (or concentration difference)
✓ Diffusion distance.

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The cells and its environment (solutions)
❖ The difference in water potential between cell and solution will determine whether
water enters or leaves out of cell by osmosis.
There are three types of solution(environment) to the cells
A. Hypertonic solution - have a lower water potential (more negative Ѱ) than the cell.
B. Hypotonic solution - have a higher water potential (less negative Ѱ) than the cell.
C. Isotonic solution - have the same water potential as the cell.
What happens when animal cells are placed in different solution?
In hypertonic solution
✓ The cells lose water by osmosis and they become shrinks
✓ Such shrink animal cell is said to be crenate and the process is called carination.
In hypotonic solution
✓ The cells gain water by osmosis and swell. The pressure will eventually burst the
cells
✓ If these animal cells are RBCS then they are said to be haemolysis, and the process
is called haemolysis.
In isotonic solution - No change on cells in such cell environment.
What happens when plant cells are placed in different solution?
A. In Hypertonic solution
✓ The cells lose water by osmosis and shrink. As a result, no pressure is exerted
by the cytoplasm of the on the wall. Such plants cells are said to be flaccid.
✓ If the cytoplasm shrinks too much, it loses contact with the cell wall. Such
plant cells are said to be Plasmolysed and the process is called plasmolysis.
B. In Hypotonic solution
✓ The cells gain water by osmosis and swell. But the cells do not burst due to the
presence of cell wall. Plant cells in this condition are said to be turgid.
✓ Turgidity is important in supporting plant cells and non-woody plant stems.
❖ If the plant is kept well-watered, the cells will remain turgid. The turgid
cells will press against each other and this pressure will keep the plant
upright.
❖ If the plant is not watered, the cells will be Plasmolysed and become
flaccid. They will no longer press against each other and the support will be
lost. The plant will wilt.
C. In Isotonic solution - No change on cells in such cell environments.

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2) ACTIVE PROCESSES:
✓ It is taken place with the expenditure of metabolic energy of cells ( ATP from
cell’s respiration).
There are three main types of active processes. these are:
A. Active transport:
• It is the movement of particles against the concentration gradient.
• It is the movement of particles from a low concentration area to higher
concentration area using carrier proteins which commonly called pumps.
• The carrier proteins that used in active transport substances across the plasma
membranes are called pumps.
❖ Na+ -pumps: carrier proteins that move sodium ions through plasma membrane.
❖ K+ - pumps: carrier proteins that move potassium ions across plasma membrane.
• It is taken place in the expenditure of ATP from cell’s respiration.
B. Endocytosis:
• Is the process of engulfing large particles by the cells through plasma
membrane. The plasma membrane surrounds the particles and form vesicles
• This process requires ATP to move the membrane around the particles and
enclosed them.
There are three forms of Endocytosis. These are:
i) Phagocytosis: is the process of engulfing large particles or even the whole
organisms by forming pseudopodia to enclose the particles and form internal
vesicles inside the cell.
❖ Pseudopodia are the extension of plasma membrane to engulf particles by cells.
ii) Pinocytosis: is the process of ingestion or engulfing of smaller scale particles by
forming small sized pseudopodia to enclose particles and form vesicles.
❖ This differs from phagocytosis with scale and form of particles which engulfed by the cells.
❖ Mostly by pinocytosis small liquid form of substances are engulfed.
iii) Exocytosis:
• Is the process of moving substances from the inside to the outside of the cell
using ATP from the cell’s respiration.
• It is the reverse of Endocytosis which is very important to :
✓ Secrete enzymes and hormones from cells.
✓ Secrete modified proteins from golgi apparatus.
✓ Remove out cellular waste products.
✓ Remove indigestible remains from phagocytic vacuoles.

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Organelles and their functions in the living cells
Organelles: - are tiny cellular structures that are specialized to perform specific
functions.
1. The nucleus
✓ It is the center of cell that controls the whole activities of cellular processes.
✓ It has several components:
 The nuclear envelop is a double membrane that surrounds the nucleus. There
are many nuclear pores on the nuclear membrane that allow the passage of some
molecules between the nucleus and the cytoplasm.
 The nucleolus is an organelle within the nucleus. It is not membrane-bound
organelle.
❖ Its function is to synthesis the components of ribosome’s, which then pass
through the nuclear pores into the cytoplasm.
 Chromatin consists of DNA molecules bound with proteins called histones.
❖ For most of the cell cycle, the chromatin fires are loosely dispersed
throughout the nucleus.
❖ Just before a cell is about to divide, the chromatin condenses into distinct,
recognizable structures called chromosomes.
2. Mitochondria: are the sites of most of the reactions of aerobic respiration.
✓ They are surrounded by two membranes, double- membrane bounded organelle.
✓ The inner membrane is folded into cristae to increase the available surface area.
✓ Some of the reactions of aerobic respiration (Krebs cycle) take place in the fluid
matrix.
✓ The folded inner membrane (cristae) provides a large surface area for the electron-
transport system, which produces most of the ATP in the mitochondria.
3. Ribosome: are the sites of protein synthesis.
✓ They can be found free in the cytoplasm, but are also bounded to the membrane
system of the endoplasmic reticulum and forming rough endoplasmic reticulum.
✓ Each ribosome comprises two subunits that are made from RNA and protein.
✓ These subunits (RNA and proteins) are manufactured in the nucleolus.
4. Endoplasmic reticulum(ER): is a membrane system found throughout the
cytoplasm of eukaryotic cells. There are two types of endoplasmic reticulum:
→ Rough ER has ribosome on its surface and is responsible for the manufacture
and transport of proteins.
➢ Protein molecules manufactured by the ribosome pass through small pores
into the lumen (inner space) of the ER.

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 ➢ Rough ER is extensive in cells that manufacture a lot of protein, such as cells
that manufactures enzymes to be secreted into the lumen of the intestine.
→ Smooth ER has no ribosome on its surface.
➢ It is concerned with the synthesis of lipids.
➢ It also synthesizes steroid substances.
➢ It is also associated with carbohydrate metabolism and detoxification.
5. Golgi apparatus (or Golgi body): is consists of a number of flattened membrane
bounded sacs in which many proteins are modified.
✓ Proteins may be converted into glycoproteins, for example. Many of the
modifications added in the Golgi apparatus act as a kind of ‘tag’, which determine
the final destination of the molecule.
✓ Think of Golgi apparatus as a cellular post office that labels and then distributes
molecule!
✓ Many of the modified molecules are released from the Golgi apparatus in vesicles
to be carried to other parts of the cell or to the plasma membrane to pass out of the
cell by exocytosis. Some vesicles form the lysosomes.
6. Lysosomes: have no specialized internal structure and are surrounded by a single
membrane.
✓ They are formed in the Golgi apparatus and contain digestive enzymes that break
down cellular waste and debris.
✓ Lysosomes are particularly abundant in phagocytic white blood cells. Here,
enzymes from the lysosomes digest foreign cells that have been engulfed.
7. Peroxisomes: are small, membrane bounded organelles which contain digestive
enzymes for breaking down toxic materials in cell.
Organelles found in plant cells
1. Cell wall
✓ Is the crisscross arrangement of cellulose fibers in the cell wall gives it both
strength and elasticity. Because there are large ‘gaps’ (on a molecular scale)
between the fibers, the cell wall is freely permeable.
2. Vacuole: The vacuole in a plant cell is a fluid-filled sac that stores a range of
solutes.
✓ It is also important in maintaining the turgidity or turgor of a cell.
✓ When the vacuole is full of liquid (mainly water), it exerts pressure on the
cytoplasm and then on the cell wall.
✓ If the vacuole loses water by osmosis, the pressure reduces and turgor is lost. The
cell becomes flaccid.

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 3. Chloroplasts: are organelles surrounded by two membranes ( they double-
membrane bounded organelles) where photosynthesis process is taken place.
✓ There are two main regions in chloroplasts that are linked to the stages of
photosynthesis.
 A Membranous regions called grana (each of this is a stack of thylakoids)
where the light-dependent reactions occur.
 A fluid stroma – where the light-independent reactions occur.

How organelles are separated from the cell sample?

✓ These have been possible because of a technique called cell fractionation.
✓ This technique is based on the fact that the various masses of organelles and
depend on their size.
✓ When a mixture of organelles is spun in a centrifuge, the various types of organelle
settle out at different speeds of spinning.
✓ The large nucleus requires relatively low centrifuge speed to settle out; the much
smaller ribosome requires a much higher speed to make it out.
✓ The cell sample is stored in a suspension that is:
• Buffered – neutral PH that prevents damage to the structure of proteins
including enzymes.
• Isotonic - prevents osmotic gain or lose by the organelles; getting to much
water could rapture the organelles.
• Cool – to reduce the overall activity of enzymes released later in the
procedure.
✓ The cells are homogenized in a blender and filtered to remove debris.
✓ The homogenized sample is placed in an ultracentrifuge and spun at low speed,
and then the nuclei settle out first and separated.
✓ The supernatant (the suspension containing the remaining organelles) is spun at a
higher speed and then chloroplast settles out if plant tissue is used.
✓ The supernatant is spun at higher speed still and mitochondria, lysosomes,
peroxisomes, fragments of plasma membrane and endoplasmic reticulum are
settling out respectively.
✓ The process is repeated at over higher speeds and finally the smallest ribosome
organelles settle out and separated.

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