BIOLOGY

LABORATORY REPORT

Name: Nurseiit Alibek

Tutor: Dr. John Carey

Group: B

Lab partner’s name: Ismagulov Galym

Title: Observing Mitosis

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Introduction
Every cell goes through a process of growing and division. This process is called the cell
cycle which is divided into three main stages: interphase, cytokinesis and mitosis (Kent 2000,
74-75). During the interphase, the cell grows and prepares itself for undergoing the mitosis.
The duration of interphase is about 90% of the cell cycle. The next stage is mitosis. Mitosis is
the division of the nucleus (Kent 2000, 74). It produces two exactly the same cells with the
same number of chromosomes, as the original cell. Mitosis involves four major phases:
prophase, metaphase, anaphase and telophase (Kent 2000, 76-77). Mitosis (Stein Carter 2010)
bеgins with prophase where strands of chromosomes become visible and the nucleoli
disappears. Due to the condensation of the chromosomes, the DNA is going to be inactive,
enabling them to move easily. After that, the cell begins metaphase where centromeres line up
on the equator. As a result, spindle apparatus is formed. Centrosomes are located at the
opposite poles of the cell. Anaphase starts, when daughter chromosomes are separated and
moved to opposite poles. The final stage is telophase. In telophase, the chromosomes cluster
at the two poles. As a consequence, the nuclear membrane is re-formed. In addition, the cell is
in the interphase before the mitosis and mitosis is followed by cytokinesis. Cytokinesis can
appear during telophase, after telophase or might not result at all (Kent 2000, 77).

Hypothesis:
It is very difficult to look at these stages under a light microscope, because they occur very
fast. Furthermore, the shortest stage of mitosis is anaphase and the longest phase is interphase.

Aims & Objectives:

 To understand the process of different stages of mitosis.
 To visually identify the stages of mitosis in a garlic root tip.
 To calculate the mitotic index and the percentage of the cells in every stage of mitosis.

Safety precautions:
During this practical, hydrochloric acid and orcein ethanoic stain were used. They are both
corrosive and irritant. We should wear goggles and gloves to avoid contact of any reagent
with eyes and skin. Moreover, acetic alcohol was used. It is corrosive and highly flammable.
We should wear goggles, lab.coat and gloves to avoid skin contact.

To avoid emergency situations every student should follow this instuctions:

 All procedures with adding reagents MUST be done in the fume cupboards.
 Be ACCURATE when using hydrochloric acid, orcein ethanoic stain and acetic
alcohol.
 ALWAYS wear goggles, laboratory coat and gloves.
 If contact does occur with any reagent in this practical, then, WASH that area with
water.

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Materials and Methods
Materials:

 Garlic roots  2 Pipettes or small measuring

 Water bath at 60 0C cylinders
 1M hydrochloric acid  2 Watch glasses or small sample
 Orcein ethanoic stain (acetic glasses
orcein)  Microscope slides and coverslips
 Acetic alcohol (ethanoic alcohol)  Pair of fine forceps
 Ice cold distilled water  Filter paper or soft tissue paper
 Test tube  Scissors

Preparation of specimen:

To start our practical part, a test tube with 2 cm3 of 1M hydrochloric acid was placed into a
water bath at 60 oC. Then, 1 cm of some garlic root tips, which were white and with rounded
end, were cut off. These root tips were put into a 2 cm3 of acetic alcohol in a watch glass for
10 minutes. After that, the root tips were placed in a second watch glass with 5 cm3 of ice-
cold water and was left for 4-5 minutes. In the next stage, root tips were put into hydrochloric
acid, which was in the water bath before, for 5 minutes. Again, these root tips were placed
into 2 cm3 of acetic alcohol for 10 minutes. Then, one of that root tips was transferred to a
microscope slide and 2-3 cm were cut from the root tip. One drop of acetic orcein stain was
added to that root tip and was left for 2 minutes. The slide was covered with a coverslip. Final
slide was viewed under the light microscope at 400x magnification. All observations were
written into the logbook.

Results
Table 1. Description of mitotic phases.

Phase Observation (image) Explanation

The nucleus replicates its DNA.

Interphase Prepares itself for cell division.
Few nucleoli become visible.

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 Chromosomes become visible and
nucleoli disappears. Also, centrioles
Prophase start to move to the poles.
The longest phase of mitosis.

Centromeres line up on the equator

Metaphase and centrosomes are located at
opposite poles of the cell.

Anaphase Chromatids go to the opposite poles.

The shortest phase of mitosis.

Telophase Nuclear membrane re-forms.

Table 2. The number of cells in each stage. (400x magnification)

Telophase
Interphase Prophase Metaphase Anaphase (might be Total
cytokinesis)

131 15 5 1 4 156
Table 3. The percentage of cells in each stage.
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 Telophase
Interphase Prophase Metaphase Anaphase (might be Total
cytokinesis)

83.97% 9.61% 3.21% 0.64% 2.57% 100%

Calculation of percentage:

w(interphase)= 131/ 156 x 100 = 83.97%

w(prophase)= 15/ 156 x 100 = 9.61%

w(metaphase)= 5/ 156 x 100 = 3.21%

w(anaphase)= 1/ 156 x 100 = 0.64%

w(telophase)= 4/ 156 x 100 = 2.57%

Diagram 1. Ranking of each stage.

Rank of each stage

100
90
80
70
60 Percentage of the cells
50
40
30
20
10
0
Interphase Prophase Metaphase Telophase Anaphase

Calculation of mitotic index.

Formula of mitotic index:

number of cells containing visible c h romosomes

Mitotic index =
total number of cells ∈t h e field of view

Mitotic index = 25/ 156 = 0.16

Discussion:
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Reliability and Validity:

I do not think that our results can be reliable, because I with my partner did this experiment
only one time. That’s why the results can not be reliable, but if we did this practical for
several times, they could be reliable. However, we have done a very good preparation of
specimen. As a result, we got a clear image of the stages of cell cycle under a light
microscope.
The results are valid and every person can repeat this practical. It is valid due to the repeatable
observations, which was made with appropriate apparatus. All measurements were done and
experimental procedure was done accurately too.

The main goal of this laboratory work was to visually identify the stages of mitosis in the
garlic root tip. During this practical, ice cold water was used to make the cells nonliving,
because nonliving cells do not actively divide. As a result, we can observe the stages of
mitosis without any fear of that the cells will move into the another phase, when you record
your data. In addition, orcein ethanoic stain was used to visually identify chromosomes
(Wikimedia Foundation, Inc.. 2013).

Firstly, I think that my hypothesis was correct and this experiment proved it. According to the
books about mitosis, we know that the longest phase of cell cycle is interphase (Kent 2000,
74). However, the shortest phase of mitosis is anaphase. We can compare our results with this
information from the book. In our experiment, the majority of cells were in the interphase and
the smallest amount of cells were in the anaphase. As I mentioned before in the reliability and
validity section our results are not reliable, because we did this experiment only one time. I
think we need more than 3 hours for this practical to get more reliable results, because a lot of
time is required to prepare very good specimen.

Another noticeable feature is an error in calculation, because cytokinesis occurs after or

before the telophase, or might not result at all. It was a little bit confusing to determine
whether the cell was in the telophase or in the cytokinesis.

Moreover, in this practical we should cut the root tips with rounded end, but it is too difficult
to know whether you are the first who cut off this root tip. The main problem is that you can
be the second who cut the same root tip. As a consequence, you will not get the root with
rounded end and you will fail your practical because you will not observe the area of active
cell divission. To avoid these kind of situations, I think that every student should be provided
with the garlic root tips on the table in front of him.

Conclusion
To sum up, during this laboratory work the stages of mitosis in the garlic root tip was clearly
observed and some suggestions for improving this practical was written. The number of cells
in the interphase (longest stage) and anaphase (shortest phase) were 131 and 1 respectively.

Reference list

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1) Kent, Michael .Advanced Biology: ''Mitosis''. New York: Oxford University Press, 2000.
pp. 74-77.

2) Stein Carter, Jan. 2010. “Mitosis”. Last modified October 2010.

http://biology.clc.uc.edu/courses/bio104/mitosis.htm

3) Wikimedia Foundation, Inc.. 2013. “Mitosis”. Last modified October 2013.

http://en.wikipedia.org/wiki/Mitosis

4) Carboni, Giorgio. 2010. “Mitosis in the root tips of garlic and onion”. Last modified April
2010. http://www.funsci.com/fun3_en/mitosis/garlic.htm

5) Wikimedia Foundation, Inc.. 2013. “Orcein”. Last modified February 2013.

http://en.wikipedia.org/wiki/Orcein

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