UDBB1104 CELL BIOLOGY

FACULTY OF SCIENCE
UNIVERSITI TUNKU ABDUL RAHMAN
Jalan Universiti, Bandar Baru Barat, 31900 Kampar, Perak.

LABORATORY PRACTICAL REPORT

TITLE Cell division: Mitosis and Meiosis

PREPARED BY NAME STUDENT ID.

1) Eswaran 1) 1401660
2) Kung Chen Han 2) 1401490
3) Lim Eng Hang 3) 1404630
4) Bong Willie 4) 1304264

DATE 8 August 2014

LECTURER DR EE KAH YAW

CRITERIA SCORE
Introduction (10%)
Materials & Methods (15%)
Results (30%)
Discussion (30%)
Conclusion (10%)
References (5%)
TOTAL (100%)

Notes:
TITLE

Cell division: Mitosis and Meiosis

OBJECTIVES

1. To be able to identify different stages of mitosis in a given sample of onion root tip.
2. To be able to identify different stages of meiosis in a prepared sample.
3. To be able to compare and contrast stages of mitosis and meiosis
4. To be able to describe activities in stages of mitosis and meiosis
5. Determine when cytokinesis occurs and the importance of this stage in cell division

INTRODUCTION

Biological concept of reproduction includes more than just the birth of new organisms.
Reproduction actually occurs much more often in cellular level. When a cell undergoes reproduction
or cell division, two daughter cells that result are genetically identical to each other and to their
parent cell. Before the parent cells split into two daughter cells, they duplicate its chromosomes and
the structure that contain cells DNA. During the division process, a set of chromosomes and
structure of cells DNA is divided to each daughter cell. As a result, two daughter cells are
genetically identical to each other and their parental cell. (J. Reece, M. Taylor , E. Simon , J.
Dickey;, 2014)

The process of cell division is a key component of the cell cycle. Most of the cell cycle is
spent in interphase. It is an order sequence of events that extends from a cell is first formed from a
dividing parent cell until its own division into two cells. The cell cycle consist two main stages:
interphase (growing phase) and mitosis phase (cell division). Interphase can divide into three sub
phases: G1 phase, S phase and G2 phase. S phase is known as DNA replication. Initially,
chromosome in cell is single, after replication, number of chromosome is double. (J. Reece, M.
Taylor , E. Simon , J. Dickey;, 2014)

After interphase is mitosis phase, also known as M phase. Cells spent only about 10% of the
total time of cell cycle. Mitosis can divide into two sub phases: mitotic phase and cytokinesis. In
mitotic phase, the nucleus and its contents are divided and distributed evenly, forming two daughter
cells. During cytokinesis, cytoplasm is divided in two. This process occurs before mitosis ends. The
combination of mitotic phase and cytokinesis produce two genetically identical daughter cells with
each consist of one set of chromosome in a nucleus, complete function of organelles and bounded by
plasma membrane. (J. Reece, M. Taylor , E. Simon , J. Dickey;, 2014)

Mitosis is a continuum process and it can divide into four main stages: prophase, metaphase,
anaphase, and telophase. In prophase, chromatin fibers are tightly coiled and folded. Each duplicated
chromosome appears as two identical sister chromatids joined together at centromere. Nucleus

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membrane is going disappear in this stage. In metaphase, chromosomes align in the center of cell.
Centrosomes are at opposite poles and spindle fiber is fully formed, attaching to the centromere.
Anaphase begins when the two centromeres of each chromosome come apart, separating the sister
chromatids. This separation is cause by the contraction of the spindle fiber. In telophase, daughter
nuclei appear at two poles of the cell as nuclear envelopes form around the chromosomes. At the end
of telophase, the chromatin fibers uncoil and the spindle fiber disappear. Cytokinesis is the division
of the cytoplasm, usually occurs along with telophase. Two daughter cells completely separated after
the end of mitosis. (J. Reece, M. Taylor , E. Simon , J. Dickey;, 2014)

In human, a typical body cell (somatic cell) has 46 chromosomes, that is 2n = 46. The
exceptions are egg and sperm cell, known as gametes. The remaining 22 pairs of chromosomes
found in both male and female are same, known as autosome. Each chromosomes consist of two
sister chromatids, arrange in pairs. The two chromosomes of such a matching pair are called
homologous chromosomes as they carry genes controlling the same inherited characteristic. A gene
that determines whether a person has freckles is located at a particular place called locus. The
difference in chromosomes between male and female are the sex chromosome. Male has an X
chromosome and Y chromosome. Female has two X chromosomes. (J. Reece, M. Taylor , E. Simon ,
J. Dickey;, 2014)

Meiosis is a type of cell division that produces haploid (n) gametes in diploid (2n) organism.
Two haploid gametes may then combine via fertilization to restore the diploid state in the zygote. If
it were not for meiosis, each generation would have twice as much genetic material as the generation
before. Meiosis is like mitosis as they precede the duplication of chromosome. This single
duplication is followed by two consecutives cell division, known as meiosis I and meiosis II. The
duplication of chromosomes is followed by two divisions. Each of four daughter cells is produced
from meiosis, which has only a haploid set of chromosomes. Meiosis I can be categorized into four
stages: prophase I, metaphase I, anaphase I, and telophase I. Meiosis II can be categorized into four
stages: prophase II, metaphase II, anaphase II, and telophase II. (J. Reece, M. Taylor , E. Simon , J.
Dickey;, 2014)

Most of the meiosis reproduction activities are same as mitosis, such as centromeres move to
opposite poles, forming of spindle fiber, and disappear of nucleus membrane. In prophase I,
chromosomes condense and coil up, become visible. In homologous chromosomes, two sister
chromatids come together as pair and this part is known as synapsis. The resulting structure
consisting of four chromatids is called tetrad. During synapsis, chromatids of homologous
chromosomes exchange certain segment, this process is called crossing over. Crossing over can
make and important contribution to the genetic variability resulting from sexual reproduction.

At metaphase I, chromosome tetrads are aligned on the metaphase plate. In each tetrad, the
spindle fiber attaches to one of the homologous chromosomes from one pole of the cell, and the other
pair of homologous chromosomes is attached by the opposite pole of the cell. With this arrangement,
the homologous of each tetrad are poised to move toward opposite pole of the cell. Like anaphase of
mitosis, anaphase I of meiosis is noticeable by migration of chromosomes towards the opposite poles
of the cell. In contrast to mitosis, the sister chromatids remain attached at their centromere, only the
tetrads split up. The still-double chromosomes moving toward each spindle pole.

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 In telophase I, the chromosomes arrive at the poles of the cell. Each chromosome is still in
duplicate form. When the chromosomes finish their journey, each pole of the cell has a haploid
chromosome set (two sister chromatids). Usually, cytokinesis occurs along with telophase I and two
daughter cells are formed. In some organisms, the chromosomes in daughter cells uncoil and nucleus
membrane reform, and there is an interphase begin before meiosis II. But some organisms
immediately proceed to second meiotic division after the first meiotic division is completed. In these
both cases, no chromosomes duplication occurs between telophase I and on the beginning of meiosis
II. (J. Reece, M. Taylor , E. Simon , J. Dickey;, 2014)

Meiosis II is also basically the same as mitosis. The main difference is that meiosis II begin
with a haploid cell. In prophase II, spindle fiber forms and moves the chromosomes toward the
center of the cell. In metaphase II, chromosomes are aligned on the metaphase plate as they are in
mitosis, with the kinetochores of the chromatids of each chromosome pointing towards opposite
poles. In anaphase II, the centromere of sister chromatids are separated. Each pair of sister
chromatids move toward to opposite poles and form individual daughter chromosome in each cells.
In telophase II, nuclei form at the cell poles and cytokinesis occurs at the same time. There are now
four haploid daughter cells. (J. Reece, M. Taylor , E. Simon , J. Dickey;, 2014)

METHODOLOGY

Mitosis

1. The tip of a root was plugged out from a provided onion sample using a two forceps.
2. The root tip was then flattened by pressing it in between two slides. The two slides were not
pressed too hard to ensure the root tip cells are not destroyed.
3. Acetocarmine was added to the sample and left to dry for a while, the slide was heated using
an ethanol lamp to enhance the absorbance of the dye into the onion root tip cells.
4. The slide with the sample was then rinsed with a little distilled water.
5. The slide was dried using laboratory paper towels.
6. Upon drying, the slide was observed under a compound microscope. The cells were first
observed under low magnification and the highest magnification (oil immersion)
7. Various stages of mitosis in the sample was identified and pictures were taken.

Meiosis

1. A prepared slide of Meiosis 1 and Meiosis 2 was observed under a compound microscope.
2. Both of the slides for Meiosis 1 and Meiosis 2 was first observed under low magnification
and the highest magnification (oil immersion)
3. Various stages in Meiosis 1 and Meiosis 2 was identified and pictures were taken.

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RESULTS
MITOSIS
Microscopic Image Sketch Description
Prophase Chromatin fibers become tightly coiled to
form condensed chromosomes. The nucleoli
disappears. Chromosomes now appear to be
identical sister chromatids joined at the
centromeres. Mitotic spindle fibers which
consist of centromeres ant the microtubules
start to form. Whereby the extended version of
these spindle fibers are known as asters.

Metaphase Centrosomes reach the opposite poles of the

cell. Chromosomes have been aligned at the
centre of the cell metaphase plate. The
kinetochores of individual sister chromatids
for each chromosome are attached to
kinetochore microtubules from opposed poles.

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 Microscopic Image Sketch Description
Anaphase Two sister chromatids of each pair moves a
part as the cohesin proteins are sliced. Two
daughter chromosomes move towards
opposite poles of the cell while the kinetochore
microtubules attached to centromeres of these
daughter chromosomes begin to shorten. Both
poles of the cell now have equal and complete
set of chromosomes.

Telophase Nuclear envelope reappears, the chromosomes

are not as much of condensed in this stage.
Spindle fibers disintegrate. Mitotic division is
now complete and two genetically identical
and independent daughter cells are formed

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MEIOSIS
Microscopic Image Sketch Description
Prophase I In this stage, nuclear membrane starts to
disintegrate and homologous chromosomes
are formed in pairs. Chromatid of homologous
chromosomes exchange segment in a process
called crossing over. They change a specific
segment of genetic information with each
other. By the way, centrosomes are moving to
the opposite pole and spindle fiber forms.

Metaphase I In this stage, chromosome tetrads align on the

metaphase plate and centrosomes are at the
opposite poles. Each spindle fiber attach to one
kinetochore at the centromere of chromosome.
They are now each linked to pole of the cell.

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 Microscopic Image Sketch Description
Anaphase I In this stage, homologous chromosomes are
divided by the pulling of spindle fiber. They
are moving to each pole of cell. However, the
sister chromatids making up the chromosome
double are still remain attached at the
centromere.

Telophase I In this phase, chromosomes arrive at the poles

of the cell. Spindle fiber disappear and nucleus
membrane reform. The cell now has haploid
chromosomes. Each of them consists of two
sister chromatids. Cleavage furrow form
between both the poles and cytokinesis occurs.

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 Microscopic Image Sketch Description
Prophase II In this stage, nuclear membrane break up
while the centrosomes form the spindle fiber.
Spindle fiber moves to opposite pole of the
cell. Chromosomes do not replicate in this
phase of meiosis. Chromosomes are beginning
migrating to the cells equator.

Metaphase II In this stage, chromosomes line up at the

metaphase II plate at the cells centre. The
kinetochore of the sister chromatid is pointing
toward opposite poles.

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 Microscopic Image Sketch Description
Anaphase II In this stage, sister chromatids are separated
and begin moving to opposite poles of the cell.
Once the sister chromatid is separated from
another, they are considered a chromosome.
Two cell poles move further apart to elongate
the cell, then forming two daughter cells.

Telophase II In this phase, distinct nuclei form at the

opposite poles and cytokinesis occurs. At the
end of meiosis II, four daughter cells are
formed. Each cell has half number of
chromosomes (haploid) as the original parent
cell.

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DISCUSSION

In this experiment, we used the onion root tips as the sample for observing mitosis. Onion roots
are easy to grow in large numbers. Also, the cells at the tips of roots are actively dividing, and thus
many cells on that particular region will be appeared in stages of mitosis. There are three cellular
regions near the tip of an onion roots. The front most region is protective root cap which contains cells
that cover and protect the growth region as the root growth through the soil on ground. The middle
part is the region where it contains the highest percentage of cells actively dividing. After cell division,
the cells continue to grow and are increasing in size in the region of cell elongation (Rainis 2010, p.
99).

The onion root tips sample was prepared by squashing the small portion of fresh root tip from
prepared onion in order to allow them to be flattened and become thinner on the microscopic slide, so
that the chromosomes of individual cells can be observed under microscope. Also, thinner sample
prepared could minimize the cells from overlapping on the slide and hence allow more lights to
penetrate through the sample during microscopy, resulting in a better and clearer image produced. At
the same time, this can prevent us from confusing the correctness of result obtained affected by the
multilayer or overlapping of cells being viewed on the microscope. The ideal thickness of microscopy
slide would be one-cell-thick.

The sample was then stained by using acetocarmine to enhance the contrast of microscopy
image produced. Acetocarmine is actually stains the plant chromosomes. The onion root tip sample
was transparent and nearly colourless before being stained by acetocarmine and the sample was very
hard to be observed under a compound light microscope. This is because transparent sample has low
contrast to the microscope, where the difference in appearance between the sample and its background
is very minor, hence the image formed is nearly invisible. The prepared onion root tip was then stained
by few drops of acetocarmine. We only wish to stain the chromosomes but not the whole cells, so the
remaining acetocarmine was washed away by running tap water. After the washing procedure, the
stained chromosomes will remain dark red in colour while the others will return to colourless since
they were unstained by acetocarmine.

For the acetocarmine stain to completely fix into the cells, it takes time up between ten to
twenty minutes which is too long for us to wait. Therefore, we warmed it gently for few times by using
an ethanol lamp. The heat produced by the flame from ethanol lamp forces the acetocarmine to
penetrate into cells in order to stain the chromosomes in faster rate. But not to heat the slide for a long
time because it will boil the acetocarmine and destroyed the root cells to be observed under
microscope. Besides, heating for long time will also causes the acetocarmine to overstrain the cells,
resulting in difficulty to observe the sample under microscope. Also, the high temperature will cause
the acetocarmine to evaporate and eventually dry very fast and stick on the slide, causing problem in
washing the slide with running tap water to remove excessive acetocarmine stain.

When we observed the mitosis of onion cells with self-prepared slide, we founded there was
very less events of mitosis can be seen and it is hard to only find a single stage of mitosis. We only
managed to observe the prophase of mitosis while the others could not be observed. On the other hand,
the observation of commercial slide was very effective, where we can find all the four stages occurs

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in mitosis just in one sample. But, the same problem is only the prophases were found abundant in
number, but the other stages of mitosis were far less in number. This is because every stage has
different duration for completion, some stage takes longer time and some stage completes very fast.
The prophase is the longest phase in mitotic division. Cells spend about 14 percent of the cell cycle in
prophase, which is the second longest phase in the complete cell cycle next to the interphase.
(McKinley n.d).

For meiosis, we used the multi-stained commercial slide provided of cross sectional of lily
anther to study the meiosis stages. Anther is the male part of the reproductive system in a flower plant;
most of the flowers produce male spores, which are also known as microspores, in the anther. The
microspore is then develops and manufactures into pollen grains in the anther. Young anther was
chosen to study because the pollen mother cells on the inner wall of the anther are still actively dividing
to produce haploid gametes, thus many cells on that particular region will be appeared in stages of
meiosis (Nuffield Foundation 2011).

The non-dividing phase which known as interphase has sometimes been called a resting stage
as no visible change could be observed under the microscope. This is totally inaccurate because the
cell is still carrying on life processes just not actively dividing. In all the G1 phase, S phase and the G2
of the interphase in cell cycle, the cell is not resting, but is actively involved in normal cellular activities
such as protein synthesis and cell growth. It also undergoes DNA replication and preparation for cell
division. Hence, the interphase is better to be known as a preparative stage rather than the resting stage
(Sreekrishna 2006, p. 99).

The time taken or the speed of cell division is not the same for all the cells, eukaryotic cells are
dividing at different rates. Single-celled eukaryotes such as amoeba, euglena and paramecium, divide
rapidly when the environment conditions are optimum to their behavior. The environment conditions
stated would be the pH value, temperature, and concentration of oxygen, nutrients, and sometimes the
presence of sunlight. On the other hand, in multicellular organism, different types of cells have
different division behaviuors (Kratz 2009).

The epithelial cells on our skin and mucous membranes are dividing all the time because they
will be shed from the body constantly, so they need to divide very frequently to replace the lost cells.
Besides, there are some cells only divide when there is any dividing signal. For example, the liver cells
do not divide normally, but may be triggered to divide if the organ is damage. Moreover, there are
some specialized cells such as neurons and red blood cells, lose their ability to replicate when they
mature. For both, basically, they were divided specially from a stem cell. Red blood cells really cannot
replicate as they have no nucleus, but are continually made from adult stem cells in bone marrow.
Neurons, on the other hand, were made from embryonic stem cells and once dead are gone forever.
For an example, if you have an injury that involves the nerve damage in the spine, the nerves cant be
repaired (Kratz 2009).

The reason for specialized cells to not replicate is that they lack of ability to go back into G1
phase after cytokinesis. The cell cycle was terminated by G0 phase. G0 is a special phase for specialized
cells which comes after mitotic phase. Once in G0, the cell either stops to divide and exits the cell
cycles, or terminally differentiated, or proceeds to G1 phase to begin a fresh cycle. Hence, both neurons

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and red blood cells are in G0 phase, a non-dividing state. These cells undergo permanent terminal
differentiation. But, majority of cells are resting in G0 for a time, but can easily slip into G1 and re-
enter the cell cycle when they need to replicate, induced by specific stimulation (Bnfalvi 2009, p.
106).

During cytokinesis process, the cell will be divided into two daughter cells. In plant cells, a cell
plate will be formed from many small vesicles moved by phragmoplast to the cell's midplane that
gradually fuse to form a large, flat vesicle that grows in diameter and change into cell membrane and
then surrounded with cell wall (Kendrew 2009, p. 247). After telophase, the content of cell was
duplicated. The two nucleuses and two set of chromosomes will be formed at the opposite poles of the
cell before cytokinesis process takes place (Ram 2010, p. 17). Hence, the cytokinesis had to occur in
the cell's midplane, right between both poles, in order to equally divide the cells content into two
distinct daughter cells. Cytokinesis that takes place at the midplane ensures that both daughter cells
end up with same size and consist of evenly distributed cellular components. This probably maximizes
the likelihood of survival and function of each daughter cell.

If a cell only underwent mitosis without cytokinesis, the cell would just end up being looks like
one big cell remains on telophase of mitosis. Inside of this cell, there would be a double set of
chromosomes because it wouldn't have split into two separate individual identical daughter cells, thus
forming a large binucleate cell where a single cell contains two nucleuses. Additionally, if the cell
continues to replicate, this will cause to the formation of multinucleated cell in which a single cell with
many nucleuses. These do exist in nature and the multinucleated cell is known as Langhans cell, which
is a large cell containing many nuclei.

CONCLUSION

In this experiment we were able to observe various stages of cell division in both mitosis and
meiosis. However we were unable to observe all the cell division stages in a given sample due to the
factor that every stage in cell division requires different amount of time to occur. Some stages of cell
division require longer time to take place while other stages seem to require a shorter time. This has a
close relation to the abundance of a particular stage of cell division in sample. We believe that the cell
division stages that took the most time appeared more in number when observed. We were also able
to outline the differences in cell division between mitosis and meiosis. The crucial role of cytokinesis
in cell division was identified and studied

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REFERENCE

J. Reece, M. Taylor, E. Simon, J. Dickey, 2014, Campbell Biology, 7th Edn, Pearson, USA.

Rainis, K.G. 2010, Cell and Microbe Science Fair Projects, Enslow Publishers, USA.

Nuffield Foundation, 2011, Preparing An Anther Squash. Retrieved on 4th August 2014, from
http://www.nuffieldfoundation.org/practical-biology/preparing-anther-squash

McKinley, n.d, Unit 4: Cell Cycle, Mitosis, Meiosis. Retrieved on 2nd August 2014, from
https://www.msu.edu/~mckinl29/unitmitosis.htm

Sreekrishna, V. 2006, Comprehensive Biotechnology II: Including Cell Biology, Genetics

Microbiology and Immunology, New Age International, Delhi.

Kratz, R.F. 2009, Molecular and Cell Biology For Dummies, John Wiley & Sons, Indiana.

Kendrew, J. 2009, Encyclopedia of Molecular Biology, John Wiley & Sons, USA.

Ram, M. 2010, Fundamentals of Cytogenetics and Genetics, PHI Learning Private Limited, New
Delhi.

Bnfalvi, G. 2009, Apoptotic Chromatin Changes. Biomedical and Life Sciences SpringerLink:
Springer e-Books, Springer Science & Business Media.

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