Lab Research 7. Prokaryotic and Eukaryotic Cells

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Professor: MD. in ECh.

Miguel Alberto López Sánchez


By: Ana Pamela Salsberg Medina
Subject: Biology I
Semester: 3rd.
Group: 37 BB
Bicultural Highschool
Introduction
Cell, in biology, the basic membrane-bound unit that contains the
fundamental molecules of life and of which all living things are composed. A
single cell is often a complete organism in itself, such as a bacterium or yeast.
Other cells acquire specialized functions as they mature. These cells
cooperate with other specialized cells and become the building blocks of
large multicellular organisms, such as humans and other animals. Although
cells are much larger than atoms, they are still very small.
A cell is enclosed by a plasma membrane, which forms a selective barrier that
allows nutrients to enter and waste products to leave.
The interior of the cell is organized into many specialized compartments, or
organelles, each surrounded by a separate membrane. One major organelle,
the nucleus, contains the genetic information necessary for cell growth and
reproduction. Each cell contains only one nucleus, whereas other types of
organelles are present in multiple copies in the cellular contents, or
cytoplasm.
Organelles include mitochondria, which are responsible for the energy
transactions necessary for cell survival; lysosomes, which digest unwanted
materials within the cell; and the endoplasmic reticulum and the Golgi
apparatus, which play important roles in the internal organization of the cell
by synthesizing selected molecules and then processing, sorting, and
directing them to their proper locations.
In addition, plant cells contain chloroplasts, which are responsible for
photosynthesis, whereby the energy of sunlight is used to convert molecules
of carbon dioxide (CO2) and water (H2O) into carbohydrates.
Between all these organelles is the space in the cytoplasm called the cytosol.
The cytosol contains an organized framework of fibrous molecules that
constitute the cytoskeleton, which gives a cell its shape, enables organelles to
move within the cell, and provides a mechanism by which the cell itself can
move. The cytosol also contains more than 10,000 different kinds of
molecules that are involved in cellular biosynthesis, the process of making
large biological molecules from small ones.
Specialized organelles are a characteristic of cells of organisms known as
eukaryotes. In contrast, cells of organisms known as prokaryotes do not
contain organelles and are generally smaller than eukaryotic cells. However,
all cells share strong similarities in biochemical function.
Cells contain a special collection of molecules that are enclosed by a
membrane. These molecules give cells the ability to grow and reproduce. The
overall process of cellular reproduction occurs in two steps: cell growth and
cell division.

Prokaryotic Cells
A prokaryote is a simple, single-celled organism that lacks a nucleus and
membrane-bound organelles. But the main thing to keep in mind for now is
that prokaryotic cells are not divided up on the inside by membrane walls but
consist instead of a single open space.
Characteristics of Prokaryotic Cell:
 Prokaryotic cells have different characteristic features:
 They lack a nuclear membrane.
 Mitochondria, Golgi bodies, chloroplast, and lysosomes are absent.
 The genetic material is present on a single chromosome.
 The histone proteins, the important constituents of eukaryotic
chromosomes, are lacking in them.
 The cell wall is made up of carbohydrates and amino acids.
 The plasma membrane acts as the mitochondrial membrane carrying
respiratory enzymes.
 They divide asexually by binary fission. The sexual mode of
reproduction involves conjugation.

Structure:
A prokaryotic cell does not have a nuclear membrane. However, the genetic
material is present in a region in the cytoplasm known as the nucleoid. They
may be spherical, rod-shaped, or spiral. A prokaryotic cell structure is as
follows:
1. Capsule– It is an outer protective covering found in the bacterial cells,
in addition to the cell wall. It helps in moisture retention, protects the
cell when engulfed, and helps in the attachment of cells to nutrients
and surfaces.
2. Cell Wall– It is the outermost layer of the cell which gives shape to the
cell.
3. Cytoplasm– The cytoplasm is mainly composed of enzymes, salts, cell
organelles and is a gel-like component.
4. Cell Membrane– This layer surrounds the cytoplasm and regulates the
entry and exit of substances in the cells.
5. Pili– These are hair-like outgrowths that attach to the surface of other
bacterial cells.
6. Flagella– These are long structures in the form of a whip, that help in
the locomotion of a cell.
7. Ribosomes– These are involved in protein synthesis.
8. Plasmids– Plasmids are non-chromosomal DNA structures. These are
not involved in reproduction.
9. Nucleoid Region– It is the region in the cytoplasm where the genetic
material is present.
10.A prokaryotic cell lacks certain organelles like mitochondria,
endoplasmic reticulum, and Golgi bodies.

Eukaryotic Cells
Eukaryotic cells are defined as cells containing organized nucleus and
organelles which are enveloped by membrane-bound organelles. Examples of
eukaryotic cells are plants, animals, protists, fungi. Their genetic material is
organized in chromosomes. Golgi apparatus, Mitochondria, Ribosomes,
Nucleus are parts of Eukaryotic Cells. Let’s learn about the parts of eukaryotic
cells in detail.
Plasma Membrane: Like prokaryotes, eukaryotic cells have a plasma
membrane made up of a phospholipid bilayer with embedded proteins that
separates the internal contents of the cell from its surrounding environment.
A phospholipid is a lipid molecule composed of two fatty acid chains and a
phosphate group.

The plasma membrane regulates the passage of some substances, such as


organic molecules, ions, and water, preventing the passage of some to
maintain internal conditions, while actively bringing in or removing others.
Other compounds move passively across the membrane.

Cytoplasm: The cytoplasm comprises the contents of a cell between the


plasma membrane and the nuclear envelope. It is made up of organelles
suspended in the gel-like cytosol, the cytoskeleton, and various chemicals.
Even though the cytoplasm consists of 70 to 80 percent water, it has a semi-
solid consistency, which comes from the proteins within it. However,
proteins are not the only organic molecules found in the cytoplasm. Glucose
and other simple sugars, polysaccharides, amino acids, nucleic acids, fatty
acids, and derivatives of glycerol are found there too. Ions of sodium,
potassium, calcium, and many other elements are also dissolved in the
cytoplasm. Many metabolic reactions, including protein synthesis, take place
in the cytoplasm.

Cytoskeleton: Within the cytoplasm, there would still be ions and organic
molecules, plus a network of protein fibers that helps to maintain the shape
of the cell, secures certain organelles in specific positions, allows cytoplasm
and vesicles to move within the cell, and enables unicellular organisms to
move independently. There are three types of fibers within the cytoskeleton:
microfilaments, also known as actin filaments, intermediate filaments, and
microtubules

Flagella and Cilia: Flagella are long, hair-like structures that extend from the
plasma membrane and are used to move an entire cell. When present, the
cell has just one flagellum or a few flagella. When cilia are present, however,
they are many in number and extend along the entire surface of the plasma
membrane. They are short, hair-like structures that are used to move entire
cells or move substances along the outer surface of the cell (for example, the
cilia of cells lining the Fallopian tubes that move the ovum toward the uterus,
or cilia lining the cells of the respiratory tract that move particulate matter
toward the throat that mucus has trapped).

Endomembrane System: The endomembrane system is a group of


membranes and organelles in eukaryotic cells that work together to modify,
package, and transport lipids and proteins. It includes the nuclear envelope,
lysosomes, and vesicles, the endoplasmic reticulum and Golgi apparatus.
Although not technically within the cell, the plasma membrane is included in
the endomembrane system because, it interacts with the other
endomembranous organelles.

The Nucleus: Typically, the nucleus is the most prominent organelle in a cell.
The nucleus houses the cell’s DNA in the form of chromatin and directs the
synthesis of ribosomes and proteins.

The outermost boundary of the nucleus is the nuclear envelope. The nuclear
envelope is punctuated with pores that control the passage of ions,
molecules, and RNA between the nucleoplasm and the cytoplasm.

Chromosomes are structures within the nucleus that are made up of DNA,
the hereditary material, and proteins. This combination of DNA and proteins
is called chromatin. In eukaryotes, chromosomes are linear structures. Every
species has a specific number of chromosomes in the nucleus of its body
cells.

Chromosomes are only visible and distinguishable from one another when
the cell is getting ready to divide. When the cell is in the growth and
maintenance phases of its life cycle, the chromosomes resemble an
unwound, jumbled bunch of threads, which is the chromatin.

The endoplasmic reticulum (ER): It’s a series of interconnected membranous


tubules that collectively modify proteins and synthesize lipids. However,
these two functions are performed in separate areas of the endoplasmic
reticulum: the rough endoplasmic reticulum and the smooth endoplasmic
reticulum, respectively.

The rough endoplasmic reticulum (RER) is so named because the ribosomes


attached to its cytoplasmic surface give it a studded appearance when
viewed through an electron microscope.

The ribosomes synthesize proteins while attached to the ER, resulting in


transfer of their newly synthesized proteins into the lumen of the RER where
they undergo modifications such as folding or addition of sugars. The RER
also makes phospholipids for cell membranes.

Golgi Apparatus: Before reaching their final destination, the lipids or proteins
within the transport vesicles need to be sorted, packaged, and tagged so that
they wind up in the right place. The sorting, tagging, packaging, and
distribution of lipids and proteins take place in the Golgi apparatus (also
called the Golgi body), a series of flattened membranous sacs.

Lysosomes: In animal cells, the lysosomes are the cell’s “garbage disposal.”
Digestive enzymes within the lysosomes aid the breakdown of proteins,
polysaccharides, lipids, nucleic acids, and even worn-out organelles. In single-
celled eukaryotes, lysosomes are important for digestion of the food they
ingest and the recycling of organelles. These enzymes are active at a much
lower pH than those located in the cytoplasm. Many reactions that take place
in the cytoplasm could not occur at a low pH, thus the advantage of
compartmentalizing the eukaryotic cell into organelles is apparent.

Lysosomes also use their hydrolytic enzymes to destroy disease-causing


organisms that might enter the cell.

Vesicles and Vacuoles: They are membrane-bound sacs that function in


storage and transport. Vacuoles are somewhat larger than vesicles, and the
membrane of a vacuole does not fuse with the membranes of other cellular
components. Vesicles can fuse with other membranes within the cell system.
Additionally, enzymes within plant vacuoles can break down
macromolecules.
Ribosomes: Ribosomes are the cellular structures responsible for protein
synthesis. Because protein synthesis is essential for all cells, ribosomes are
found in practically every cell, although they are smaller in prokaryotic cells.
They are particularly abundant in immature red blood cells for the synthesis
of hemoglobin, which functions in the transport of oxygen throughout the
body.

Mitochondria: Mitochondria: They are often called the “powerhouses” or


“energy factories” of a cell because they are responsible for making
adenosine triphosphate, the cell’s main energy-carrying molecule.

Peroxisomes: Peroxisomes are small, round organelles enclosed by single


membranes. They carry out oxidation reactions that break down fatty acids
and amino acids. They also detoxify many poisons that may enter the body.
Alcohol is detoxified by peroxisomes in liver cells. A byproduct of these
oxidation reactions is hydrogen peroxide, H2O2, which is contained within
the peroxisomes to prevent the chemical from causing damage to cellular
components outside of the organelle. Hydrogen peroxide is safely broken
down by peroxisomal enzymes into water and oxygen.

Similarities and differences between prokaryotic and


eukaryotic cells
Similarities between eukaryotic and prokaryotic cells

Cell Membrane- Both eukaryotic and prokaryotic cells bear a lipid bilayer,
which is an arrangement of phospholipids and proteins that acts as a
selective barrier between the internal and external environment of the cell.

Genetic Material- Eukaryotic and prokaryotic cells both use deoxyribonucleic


acid (DNA) as the basis for their genetic information. This genetic material is
needed to regulate and inform cell function through the creation of RNA by
transcription, followed by the generation of proteins through translation.

Ribosomes- Ribosomes facilitate RNA translation and the creation of protein,


which is essential to the functioning of both eukaryotic and prokaryotic cells.
Cytoplasm- The cytoplasm is the medium in which the biochemical reactions
of the cell take place, of which the primary component is cytosol.

In eukaryotic cells, the cytoplasm comprises everything between the plasma


membrane and the nuclear envelope, including the organelles; the material
within the nucleus is termed the nucleoplasm. In prokaryotes the cytoplasm
encompasses everything within the plasma membrane, including the
cytoskeleton and genetic material.

Differences between eukaryotic and prokaryotic cells

Cell size---Eukaryotic cells are ordinarily larger (10 – 100um) than prokaryotic
cells (1 – 10um).

Cell arrangement---Eukaryotes are often multicellular whereas prokaryotes


are unicellular. There are however some exceptions –unicellular eukaryotes
include amoebas, paramecium, yeast.

True membrane bound nucleus--- Eukaryotic cells have a true nucleus bound
by a double membrane. It contains the DNA related functions of the large cell
in a smaller enclosure to ensure close proximity of materials and increased
efficiency for cellular communication and functions.

In contrast, the smaller prokaryotic cells have no nucleus. The materials are
already fairly close to each other and there is only a "nucleoid" which is the
central open region of the cell where the DNA is located.

DNA structure--- Eukaryotic DNA is linear and complexed with packaging


proteins called "histones," before organization into a number of
chromosomes

Prokaryotic DNA is circular and is neither associated with histones nor


organized into chromosomes. A prokaryotic cell is simpler and requires far
fewer genes to function than the eukaryotic cell. Therefore, it contains only
one circular DNA molecule and various smaller DNA circlets (plasmids).
Membrane-bound organelles--- Eukaryotic cells contain many membrane-
enclosed, large, complex organelles in the cytoplasm whereas prokaryotic
cells do not contain these membrane-bound organelles. This is a key
difference because it allows a high level of intracellular division of labor and
contributes to the greater complexity characteristic of eukaryotic cells.

Due to the larger size of the eukaryotic cells, confining certain cellular
process to a smaller area also increases the efficiency of functions by
improving communication and movement within the cell.

Only eukaryotes possess a membrane-bound nucleus and membrane-bound


organelles such as the mitochondria, Golgi apparatus, lysosomes,
peroxisomes, and ER.

Ribosome size--- Both eukaryotic and prokaryotic cells contain many


ribosomes; however, the ribosomes of the eukaryotic cells are larger than
prokaryotic ribosomes i.e. 80S compared to 70S.

Eukaryotic ribosomes also show more complexity than prokaryotic – they are
constructed of five kinds of ribosomal RNA and about eighty kinds of
proteins. In contrast, prokaryotic ribosomes are composed of only three
kinds of rRNA and about fifty kinds of protein.

Cytoskeleton--- This is a multicomponent system in eukaryotes composed of


microtubules, actin filaments and intermediate filaments. It is required for
maintaining cell shape, providing internal organization and mechanical
support. It is also paramount in movement and cell division.

Sexual reproduction--- Most eukaryotes undergo sexual reproduction whilst


prokaryotes reproduce asexually. Sexual reproduction in eukaryotes results
in offspring with genetic material which is a mixture of the parents’ genome
and during this process, genetic variation is generated via sexual
recombination.
On the other hand, a prokaryote will reproduce clones of itself via binary
fission and relies more on horizontal genetic transfer for variation.

Cell division--- This occurs by mitosis for eukaryotic cells and binary fission for
prokaryotic cells.

Eukaryotic cells undergo mitosis then cytokinesis. This involves numerous


stages - the nuclear membrane disintegrates then the chromosomes are
sorted and separated to ensure that each daughter cell receives two sets (a
diploid number) of chromosomes. Following this, the cytoplasm divides to
form two genetically identical daughter cells i.e. cytokinesis.

In contrast, prokaryotes undergo a simpler process of binary fission. This is


faster than mitosis and involves DNA (nucleoid) replication, chromosomal
segregation, and ultimately cell separation into two daughter cells genetically
identical to the parent cell. Unlike mitosis, this process does not involve the
nuclear envelope and centromere and spindle formation.

Microorganisms that can be found in yogurt, fermented


juices, and oral mucosal epithelium
Yogurt: Yogurt is made when heated milk is combined with bacteria,
specifically Lactobacillus bulgaricus and Streptococcus thermophilus, and left
to sit for several hours at a warm temperature (110-115°F). Additional types
of lactobacilli and bifidobacteria may be added.
Fermented Juices: The most commonly reported bacterial genera include
Acetobacter, Alicyclobacillus, Bacillus, Gluconobacter, Lactobacillus,
Leuconostoc, Zymomonas, and Zymobacter. Among yeasts Pichia, Candida,
Saccharomyces, and Rhodotorula are commonly encountered genera
responsible for spoilage of juices
Oral mucosal epithelium: Immuno-modulation, Bacterial Infection, and
Cancer Cells: There is a known association between bacterial infection and
cancer. Bacterial components are able to up-regulate immune-modulatory
receptors on cancer cells. Interactions of bacteria with tumor cells could
support malignant transformation an environment with deficient immune
regulation. The aim of this review is to present a set of molecular
mechanisms of oral epithelial cells and their reactions to a number of toxic
influences.

Staining techniques in Biology


Objectives of Staining
Improves visibility by greater contrast between the organism and the background. /
Differentiate various morphological types (by shape, size, arrangement, etc.). / Determine
the staining characteristic of organism and, at times, direct diagnosis of disease.
/Demonstrate the purity of culture. observe certain structures (flagella, capsules,
endospores, etc.).
Staining Techniques
Gram Stain:

The differential nature of the Gram stain is based on the ability of some
bacterial cells to retain a primary stain (crystal violet) by resisting a
decolorization process. Gram staining involves four steps. First cells are
stained with crystal violet, followed by the addition of a setting agent for the
stain (iodine). Then alcohol is applied, which selectively removes the stain
from only the Gram-negative cells. Finally, a secondary stain, safranin, is
added, which counterstains the decolorized cells pink.
Acid Fast Stain:

Some bacteria produce the waxy substance mycolic acid when they construct
their cell walls. Mycolic acid acts as a barrier, protecting the cells from
dehydrating, as well as from phagocytosis by immune system cells in a host.
This waxy barrier also prevents stains from penetrating the cell, which is why
the Gram stain does not work with mycobacteria such as Mycobacterium,
which are pathogens of humans and animals. For these bacteria, the acid–
fast staining technique is used.
To perform the acid-fast stain, a heat-fixed smear is flooded with the primary
stain carbol fuchsin, while the slide is heated over a steaming water bath. The
heat “melts” the waxy cell wall and permits the absorption of the dye by the
cells. Then the slide is allowed to cool, and a solution of acid and alcohol is
added as a decolorizer. Cells that are “acid-fast” because of the mycolic acid
in their cell wall resist decolorization and retain the primary stain. All other
cell types will be decolorized. Methylene blue is then used as a counterstain.
In the end, acid-fast bacteria (AFB) will be stained a bright pink color, and all
other cell types will appear blue.
Staining Methods to Highlight Specific Cell Structures
Capsule: The polysaccharide goo that surrounds some species of bacteria and
a few types of eukaryotic microbes is best visualized when the cells are
negative stained. In this method, the bacteria are first mixed with the stain,
and then a drop of the mixture is spread across the surface of a slide in the
thin film. With this method, capsules appear as a clear layer around the
bacterial cells, with the background stained dark.
Metachromatic granules or other intracytoplasmic bodies: Some bacteria
may contain storage bodies that can be stained. One example is the Gram-
positive bacilli Corynebacterium, which stores phosphate in structures called
“volutin” or metachromatic granules that are housed within the cell
membrane. Various staining methods are used to visualize intracytoplasmic
bodies in bacteria, which often provide an identification clue when observed
in cells.
Endospore Stain
Endospores are dormant forms of living bacteria and should not be confused
with reproductive spores produced by fungi. These structures are produced
by a few genera of Gram-positive bacteria, almost all bacilli, in response to
adverse environmental conditions. Two common bacteria that produce
endospores are Bacillus or Clostridium. Both live primarily in soil and as
symbionts of plants and animals and produce endospores to survive in an
environment that change rapidly and often.
The process of Endo sporulation (the formation of endospores) involves
several stages. After the bacterial cell replicates its DNA, layers of
peptidoglycan and protein are produced to surround the genetic material.
Once fully formed, the endospore is released from the cell and may sit
dormant for days, weeks, or years. When more favorable environmental
conditions prevail, endospores germinate and return to active duty as
vegetative cells.

How to prepare a bacterial smear from cultured cells for


staining and observation in microscopes
The preparation of a smear is required for many laboratory procedures,
including the Gram-stain. The purpose of making a smear is to fix the bacteria
onto the slide and to prevent the sample from being lost during a staining
procedure. A smear can be prepared from a solid or broth medium. Below
are some guidelines for preparing a smear for a Gram-stain.
1. Place one needle of solid bacterial growth or two loops
of liquid bacterial growth in the center of a clean slide.
2. If working from a solid medium, add one drop (and only one drop)
of water to your specimen with a water bottle. If using a broth
medium, do not add the water.
3. Now, with your inoculating loop, mix the specimen with the water
completely and spread the mixture out to cover about half of the
total slide area.
4. Place the slide on a slide warmer and wait for it to dry. The smear is now
ready for the staining procedure.

Use of methylene blue and gentian violet for staining


It is a cationic dye that stains cells blue because the positively charged dye is
attracted to negatively charged particles such as polyphosphates, DNAs, and
RNAs. Specimens collected from patients by swabbing are smeared onto
microscope slides and the methylene blue solution is dropped on the slide

Discussion
I was hoping to see the bacteria in the expired yogurt more than any other.
Because even though it is pasteurized milk, they basically add bacteria to it,
so if it already had bacteria, and then we give those bacteria the opportunity
to “evolve” they might be more visible. I also thought that in the oral
mucosal epithelium we’d be able to see even more bacteria and cells than in
the yogurt, because of how many bacteria we have in our mouths, which
might not be harmful to us, but they’re still abundant.

Conclusion
The structure and function of cells have now become a mix of basic research
and clinical topics. The tools have also expanded with the growth of
molecular and analytical techniques. In this Century, especially during the
past two decades, we also experienced an unparalleled rise in our
understanding of the cell, its structure and complexity, its chemical
organization, and the specific roles of its sections. This knowledge focused on
the efforts that thousands of scientists are working on throughout the world
in laboratories.
The importance of cell biology in the 21st Century is that it has helped us in
so many ways, and by understanding how cells work, cell biologists are
working in the animal, plant, and medical science. We will develop new
medicines, more effective drugs, plants with enhanced properties, and a
better understanding of how all living things function by increased
knowledge.
Eventually, it is and will be more effectively possible to produce a ‘health
forecast’ by analyzing our database of genetic and cell information. Using
this, we can take better control over our health in a preventive way.
Although, cell biology is not just about disease. It has greatly assisted the
humans in other ways, for example, they’ve helped many people find out
more about because of their DNA, which cells also happen to carry.
So yeah, cells play a massive role in life, but I guess we all already knew
that. :)

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Eukaryotic Cell Illustration Prokaryotic Cell Illustration

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