Mitosis
(feulgen reaction)
J. Ricardo; D. Fonseca; M. Fontes; R. Junceiro
Universidade de Trás-os-Montes e Alto Douro, UTAD
Quinta de Prados, 5000-801 Vila Real, Portugal
Abstract:
This experimental activity was carried out in
order to observe the different phases of mitosis
using onion root vegetative vertices, by Feulgen
reaction.
Mitosis is a process of cell division that results in
the formation of two daughter cells with the
same genetic characteristics and the same
number of chromosomes.
By observing under the microscope, we can see
that the cells in interphase appear in larger
quantity than the cells in the other phases of the
cell cycle, as this is the longest phase of the cycle
(about 15 hours). We can also observe cells in
metaphase and anaphase.
That way, mitosis is a very important process for
plant life, as it is of great importance that we
know how it occurs for future investigations.
Keywords: interphase;
3
Introduction:
Mitosis is a process of cell division that results
in the formation of two daughter cells with the
same genetic characteristics and the same
number of chromosomes. This process,
considered an equitable division, promotes
organism development, maintenance of
homeostasis, tissue renewal and asexual
reproduction.
This process of cell division is divided into
several stages, the ones we intend to observe
are prophase, metaphase, anaphase and
telophase.
This experimental activity was carried out in
order to observe the different phases of mitosis
using onion root vegetative vertices, by Feulgen
reaction.
Feulgen reaction is a chemical process with
high specificity for DNA, widely used in
cytochemistry. This reaction is based on the
chemical properties of fuchsin and the
hydrolytic action of HCl on DNA. For a long
time, this reaction has been used in DNA
quantitation methods, but today this has been
shown to be less effective in the development
of fluorophore or fluorochrome based flow
cytometry methods. For cytogenetics, this
reaction provides well-defined and clean
preparations. Despite being a somewhat more
laborious and time-consuming method when
compared to other conventional staining
techniques, chromosomes appear more visually
delimited, with the primary and secondary
constrictions more evident. Also, due to the
specificity for
DNA, other components such as RNA,
ribonucleoproteins and cytoplasm are not
stained. Slides prepared by the Feulgen method
may lessen the intensity or discolor more easily
over time on permanent slides, while those
stained with Giemsa and Hematoxylin, for
example, remain in good contrast for much
longer.
from the slide, and material may become
attached to the slide or coverslip. If the coverslip
does not come to a halt after 24 hours,
separation with a dissecting needle should be
assisted. Immediately after this process, there is
absolute alcohol dehydration, then xylol
diaphanization and finally Canadian balm
mounting.

Materials and Methods:

Results and Discussion
Turning to the material and methods we use
root vegetative vertices of onion as it is an easy After performing all the steps of the procedure,
material to obtain, presents a merismatic zone we obtained several results which consisted of
where cell division occurs and consequently, observing the various phases of the cell cycle, as
achieve the expected results. We used the slides shown in Figure 1 and 2, the existing phases of
and coverslips to observe the microscope; that cycle.
distilled water for washing. We spend different
types of alcohol, one 95% ethyl alcohol (V / V)
and one absolute, which serve to separate the
lamella and the blade, getting the material stuck
in the blade and dehydration, consecutively.
Thermo-stabilized bath, or better known as
water bath. The Carnoy fixative, which consists
of 6 parts absolute alcohol + 1 part acetic acid, is
also used to kill the material. Acetic acid 45% (V /
V) for the dissociation of vegetative vertices. We
use Schiff's reagent is a dye (fuchsin). 1N
hydrochloric acid for hydrolysis. Xylol for
diaphanization which is a conservation
Figure 1: Cell cycle phases.
technique to facilitate its study. Dissecting
needles and microscope for observation of our
preparation. Finally, we use Canada balm for
final assembly.

We start by fixating the vertices for 30 minutes

in Carnoy. Then the first wash with distilled
water occurs. We then hydrolyzed with 1N
hydrochloric acid for 3-10 (+ -6 min) minutes in a
water bath at 60 ° C. After the second wash with
distilled water, we stain with Schiff reagent for
30 minutes in the dark (as it is a photolabile
Figure 2: Representation of cell cycle phases in
substance). Next, we have the 3rd wash with
plant cells.
distilled water, dissociating the vertices in 45%
acetic acid on a glass slide and later compressing
a coverslip on the differentiated material, called
After observing the results, we can see that the
smear. Subsequently, the slide / coverslip sets
cells in interphase appear in larger quantity than

3
After observing the results, we can see that the cells
in interphase appear in larger quantity than the cells
in the other phases of the cell cycle, as this is the
longest phase of the cycle (about 15 hours). We also
noticed that the cells in metaphase are the ones that
have the highest staining due to the maximum
condensation of chromosomes in this same phase.
Anaphase, which is depicted in Figure 4, is an easy
step to identify due to the shape of the chromatids
in their separation, although there are few cells in
this phase. Prophase cells could be observed, even
though it is a little difficult, due to the formation of
condensed chromosomes. Telophase cells were also
difficult to identify because they could be confused
with anaphase cells.
It is important to emphasize the importance of using
the vegetative vertices of the onion as they have
meristems (place where mitosis occurs quite often).
Figure 3. Microscope observation.
Figure 4. Microscope observation
Figure 5. Microscope observation
Conclusions:
Thus, we conclude that in fact the merismatic zone is
a place where a lot of cell division occurs and the
most present phase was the interphase , since it is
the longest stage and its duration is 15h.Despite the
difficulty in analyzing some cells, the purpose of this
work was completed and we had the opportunity to
see the various phases of the cycle mentioned
earlier.
In short, mitosis is a very important process for plant
life, as it is of great importance that we know how it
occurs for future investigations
Bibliography:
http://
hsmonografiaroaq.capefearequality.org/mitose-
na-cylula-vegetal-49godetyco4537.html
https://pt.slideshare.net/tic2012/ciclo-celular-
45944803n