Laboratory

Mitosis,
Meiosis and
Chromosomes

2 Laboratory 8: Mitosis and Meiosis

OBJECTIVES
After completing this lab you will be able to:
1. Identify the stages of mitosis and meiosis..
2. Locate the stages of mitosis in onion root tips.
3. Find and describe the stages of meiosis in lily anthers
4. Describe polytene chromosomes is olated from fruit-fly saliva ry
glands

LAB PREPARATION
Before lab, you should do all of the following:
1.Read Chapter 12 and 13 in Campbell (8th Ed.)
2.Bring personal protective gear : lab coat, safety goggles, and
safety gloves.

INTRODUCTION
Although some genetic information is carried within organelles such
as mitochondria and chloroplasts, it is the nucleus that contains the
bulk of the cells genetic information (DNA). The process of mitosis
divides this nuclear genetic materi al equally between daughter cells
during cell division. The distribution of identical genetic information
is brought about by the longitudinal splitting of the chromosomes. A
complex spindle apparatus arranges the separation of one whole set
of identical chromosomes to each daughter cell.
The normal processes of mitosis and cell division are under tight
control within the cell. Only through the gradual emancipation of a
cell from this tight control does a cell turn cancerous. In other words,
cancer or tumor growth are the res ult of unc ontrolled cell division.
Thus, before we can understand how cancer arises, it is necessary to
understand the process of mitosis.
Meiosis, on the other hand, is nucl ear division that gives rise to
gametes in higher plants and an imals. This process provides the
opportunity for genetic recombinati on, leading to genetic variation
(diversity) from generation to ge neration. This diversity is very
important for the survival of th e species. The process of meiosis
consists of two nuclear divisi ons, and results in the number of
chromosomes in the ga metes being reduced by half. In the first
division, the reduction division, homologous chromosomes are
separated randomly between two da ughter cells, giving the haploid
chromosome number. During the second division the chromosomes
are split at the cent romere, as in m itosis, giving rise to four haploid
cells.

Laboratory 8: Mitosis and Meiosis 3

A. Mitosis in the Onion Root-tip
The onion root tip is one of the mo st widely used mate rials for the
study of the cell cycle be cause it is readily av ailable, preparation of
the dividing cells is easy, and the chromosomes are large and few in
number, hence, easier to study
than the cells of many other
organisms. Since root tips are regions of active cell division, chances
are good that in a specimen of such tissues one can find every stage
of this process. Resist the temptation to think of the various stages of
the cell cycle as a series of indi vidual stages in which one stage is
discretely followed by another; th e process is more like a movie.
However, it is easier to study the process as a seri es of individual
stages (like frames of a movie) that show the important
characteristics of each stage.
The chromosomes of many plants ar e large and eas y to see. While
there are many suitable plants for the study of mitosis, the onion has
become a standard in teaching labor atories. Root- tips, with the ir
many actively dividing cells, ar
e the preferred tissue for
examination. Preparation is simple and quick. The tissue is fixed
and softened by treatment with acid and heat, both of which serve to
break down the cell wall and membra nes. A stain is the n applied
(toluidine blue in this case), wh ich penetrates the damaged cells and
stains the chromosomes. The root-tip is finally squashed by hand in
order to spread out the cells
and the ir chromosomes for easier
viewing.
B. Meiosis in the Lily Anther
While the cells in soma tic tis sues of plan ts and a nimals r outinely
undergo mitosis, only the cells
in reproductive or gans under go
meiosis. Most animal reproducti ve cells contain numerous, small
chromosomes and hence meiosis is
dif ficult to observe. The
immature anthers of pla nts are of ten used to illustrate meiosis
because the cells are lar ge, and relatively few chromosomes are
present. Because the developing microspores (pollen grains) usually
divide synchronously, many cells in the same phase of meiosis are
often visible.
Unfortunately, the preparation of an thers for examination of meiosis
is difficult and time consuming, requiring embedding of the tissue,
staining, and the use of a microtome for sectioning. Because there is
insufficient time in this laborat ory to go through these procedures,
you will examine prepared slides obtained from a biological supply
house.
C. Giant Chromosomes of Drosophila
Without doubt, geneticists have proclaimed the fruit fly Drosophila
melanogaster the animal of choice for ge netic stud ies. While the
reasons for this choice are numer ous (e.g., they are easily cultured

4 Laboratory 8: Mitosis and Meiosis

and have a short generation time), perhaps one of the most important
reasons is the giant chromosomes that are present in the salivary
glands during the fly s larval stag e. These chr omosomes, the result
of repeated duplication of the DN A helix, are massive, and clearly
show (when properly stained) a banding pattern which geneticists
have exploited to explore the genetic basis of morphology, behavior,
etc. The c hromosomes a ttain th eir maximum s ize jus t befor e
pupation, i.e., late third instar (see Figure 7-1). They then degenerate.
The large, multi-stranded, or polyt ene, chromosomes are not limited
to the salivary glands but are also present in the gut and Malpighian
tubules of larva and the foot pads of adults. However, they reach
their largest size in the salivary glands and are most easily examined
in this tissue.

D. Heredity and Human Disorders

Meiosis occurs repeatedly in our human lifetime, as our testes or
ovaries produce gametes (sperm or eggs). Almost always, the meiotic
spindle distributes chromosomes to daughter cells without error. But
occasionally there is an accident, called a nond isjunction, in which
the me mbers of a chromosome pair fa il to sepa rate. This resu lts in
gametes with an abnormal, aneuploid number of chromosomes. Most
embryos resulting from fertilization that involves aneuploid gametes
do not proceed very far in develo pment, but are naturally aborted.
Some types of aneuploid embryos, however, can survive to birth and
beyond. Nondisjunction in meiosis ca n af fect either the autosomes
(those chromosomes not directly involved in determining sex) or the

Laboratory 8: Mitosis and Meiosis 5

sex chromosomes (the pair of chromosomes responsible for
determining the gender of an individual).
Alterations of chromosome number and structure are associated with
a number of serious human disorders, such as Down syndrome
(possessing an extra 21st chromo some) and Klinefelter syndrome (a
male posses sing an extra X chro mosome). Other human genetic
disorders are caused by alteratio ns in a gene or genes that control
specific enzymes or metabolic pathways. These genes may be
located on either the autosomes or the sex-chromosomes; the latter
disorders are said to be sex-li nked and include red-green color
blindness and hemophilia. Some
genetic disorders are more
disabling than others. Genetic a nd biochemical tests on fetal cells
and molecules, as well as examin ation of the fetus with ultrasound,
can help people make a decision
whether to keep or abort the
pregnancy.

6 Laboratory 8: Mitosis and Meiosis

LABORATORY EXPERIMENTS
You should work individually for all of todays laboratory exercises.
Experiment 1: Mitosis in the Onion Root-tip

You must wear personal protective gear (lab coat, gloves, goggles)
when working with 1N HCl, as it is a very strong acid.
PROCEDURE
1.Remove ONE ENTIRE ROOT from the base of an onion. (DO
NOT remove only the bottom port ion of the root because, if
you do, the next student will mistakenly remove the top portion
that does not contain any mitotic tissue.)
2.Cut the distal 2 mm from the ti p of the root a nd place it on a
clean microscope slide. Cover it with 2-3 drops of 1N HCl and
gently warm the slide on the sl ide warmer for 2-3 minutes at
60-70oC.
3.Remove the HCl from the slide by blotting with a small piece
of bibulous paper . Add 2-3 drops of 0.5% aqueous toluidine
blue stain and re-heat for 1-2 minutes.
4.Remove the stain by blotting wi th a small piece of bibulous
paper, and cover the root-tip with a drop of distilled water.
5.Apply a cover glass, and with a folded Kimwipe around your
thumb (to soak up the excess water) squash the root-tip. Apply
some force but DO NOT allow the cover glass to slide.
6.Examine the squashed root tip with your compound microscope
and look for the various stages of mitosis. The chromosome
number of the common onion is 2N = 16.
7.To get some idea of the length
of time that a dividing cell
spends in each of the mitotic phases, count the number of cells
you find in prophase, metaphase, anaphase, and telophase. Do
this for about 30 (or more) divi ding cells. Keep in mind that
mitosis is a continuous process, not one in which a cell jumps
from one phase to the next. As a result, you will probably find

Laboratory 8: Mitosis and Meiosis 7

some cells that don t fit nicely into any of the four mitotic
phases; classify these as best you can. If mitosis in the onion
lasts an average of 30 minutes, what is the approximate length
of each of its phases?
8.When you have finished examini ng your root tips, discard the
cover glass in the glass only box. Rinse & dry the slide, and
return it to the slide box for re-use.

MEIOSIS
In meiosis, immature or primor dial germ cells under go a reduction
from the diploid number to the haploid number of chromosomes and
become mature gametes. As a
result, meiosis maintains the
chromosome .number constant a nd provides genetic variability
because of crossing over and the
subsequent exchange of genes
between chromosomes.
Experiment 2: Meiosis in the Lily Anther
PROCEDURE
In the back of the lab there are eight empty microscopes and a set of
prepared slides showing meiosis in the lily anther. Your TA will ask
you to find an example of one
of the eight phases of meiosis:
prophase I, metaphase I, anaphase
I, telophase I, prophase II,
metaphase II, anaphase II, and telophase II. When you find the
phase, place the slide on the appropr iate empty microscope for your
classmates to look at.
Each prepared slide normally cont ains several sections through the
anther, however, not all the sections (sometimes only one!) contain

8 Laboratory 8: Mitosis and Meiosis

Laboratory 8: Mitosis and Meiosis 9

the phase of meiosis printed on the slide label. Y
ou should,
therefore, use your text to confir m your phase iden tification and not
the slide label. When looking fo r your assigned pha se, please take
only one slide at a ti me, and return that slide before obtaining
another.
Meiosis in the Lily anther will
be stu died a s it occu rs in the
development of mature pollen grains of fl owering plants. These
pollen grains give rise to male ga metes, which fuse with an egg to
produce a zygote.

Before you leave lab, make certain you can identify EACH stage of
meiosis.
Experiment 3: Examination of the Giant Chromosomes of
Drosophila Larvae
Obtain a prepared slide of polytene chromoso mes from Drosophila.
Examine th e slide first un der low power and observe how the
nucleus of each of the cells is co mposed of a coil of giant, banded
chromosomes. Choose a cell in which the coil is well spread out and
gradually increase the objective le ns magnification to 100X (oil
immersion).
Make a sketch of the banded chromosomes from one nucleus in your
notebook, showing a short segment in detail.
Be sure to completely clean off the oil from the 100X objective with
lens tissue when you are finished with your observations!

10 Laboratory 8: Mitosis and Meiosis

a. Interphase
The interphase cell, so named because early biologists thought it was in a resting phase, is
actively undergoing respiration and the synthesis of DNA, RNA, and protein in preparation
for mitosis.
b. Prophase
During prophase, the DNA, originally in long, thin strands, becomes condensed as a result of
coiling an d supercoiling. The nuclear me mbrane begins to break down, and th
e
chromosomes are distributed throughout the nucleoplasm. During prophase in the onion root
tip, the c hromosomes oft en ap pear as a co iled ma ss. Even at t his ea rly s tage, each
chromosome has doubled, though th is is difficult to see on a slide. Under very high
magnifications, it is possible to see that each chromosome is composed of two separate
strands, the sister chromatids. The two sister chromatids are identical in structure, chemistry,
and the genetic information they carry because one was replicated (copied) from the original
DNA of the other during the last S phase. The sister chromatid s are joined together at a
region of attachment called the centromere. Within this region, each chromatid contains a
disc-shaped k inetochore. Microtub ules (called kinetochor e microtubules) insert into th e
kinetochores and run from them o utward to the two po les o f the c ell. Other pola r
microtubules become organized into the spindle fibers.
c. Metaphase
During early metaphase, some of the polar microtubules break down and new attachments
are made between the kinetochore microtubules and tub ules from the opposite p ole. This
results in what appear to be rat her ai mless chromosome mov ements, aptly described as
dancing chromosomes. As metaphase progres ses, a random breaking and reattachment of
kinetochore microtubu les to the polar microt ubules of the same or opposite poles occurs
until (again randomly) the ki
netochore of on e daughter ch romatid is attached to
microtubules from one pole, and the kinetochore of the daug hter chromatid is connected to
tubules from the opposite pole. Then the polar microtubules pull in such a way th at th e
kinetochores become positioned in a region halfway between the poles. This region, which
occupies a plane near the center of the cell (and at right angles to the long axis of the spindle
fibers), is c alled the metaphase or equatorial plate. The cell is considered to have reached
metaphase when the kinetochores of a ll chromosomes have arrived at this equ atorial plate
region. At this time the centromeres d ivide in prep aration for sep aration of th e daughter
chromatids during the following stageanaphase.
d. Anaphase
The sister chromatids that make up each chromosome are separated from each other and are
pulled by the microtubules to opposite poles of the cell. As the centromeres are pulled apart,
the arms of these (now called) daughter chromosomes are passively dragged along. Thus,
anaphase in onion cells can be recognized by the twogroups of V-shaped chromosomes on
opposite sides of the cell. The sharp end of th e V is oriented toward the pole of the spindle.
Reduce the light by adjusting the diaphragm of the microscope, and try to locate any spindle
fibers near the center of the cell. They appe ar as very fine lines between the two groups of
chromosomes. Anaphase ends when the newly separated chromosomes arrive at opposite
poles of the cells.
e. Telophase
Karyokinesis is completed durin g telophase, and reor ganization of the contents of the two
daughter cells (cytokinesis) b egins. It is often difficult to distinguish late anaphase fro m
early telophase in the cells of plan ts. During telophase, however, a cell plate, the first
indication that cytokinesis is begi nning, starts to fo rm as a fine line across the center of th e
cell. When co mplete, the cell plate divides the or iginal cell into two d aughter cells. As
telophase progresses, the nuclei begin to reor ganize: the chromosomes uncoil and become
longer and thinner, the nuclear membrane reforms, and the nu cleoli reappear. Mitosis ends
with the assembly of two int erphase nuclei, each with one complete set of single-stranded
chromosomes. The daughter cells resulting from mitotic division have the same number and
kinds of chromosomes (and therefore the same genetic makeup) as the original cell.

Laboratory 8: Mitosis and Meiosis 11

12 Laboratory 8: Mitosis and Meiosis

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Laboratory 8: Mitosis and Meiosis 13

Fill in this form and submit with your Lab Summary.
QUESTIONS TO RESEARCH AND ANSWER
1.a) How many pairs o f chromosomes are contained in each human
body cell?
b)How many genes do scientists
chromosomes?

estimate ar e loc ated on those

2.a)What do most genes contain instructions for building?

b)What functions do these perform?

3.How many pairs of genes are t

color?

hought to code for human skin

4.The different forms of genes are called ______________.

5.Alleles are thought to arise by the process called
____________________.
6.While some mutations have
contributed to healthy human
diversity, most mutations are _____________.
7.Two examples of diseases caused by recessive alleles are:

8.Diseases caused by recessive genes are relatively (rare, common ,

lethal) (circle one) because they are masked by the dominant allele.
9.In contrast to diseases caused by recessive alleles, an example of a
disease caused by a dominant allele is
_________________________.

14 Laboratory 8: Mitosis and Meiosis

10.In humans, 22 of the 23 ch
_______________________.

romosome pairs are called

11.In humans, the 23rd pair

_________________________.

of chro mosomes is c

alled

12.A normal human female has 2 ______ sex chromosomes, while a

normal human male has an _____ and a ________ chromosome.
13.Most genes on the sex chromo somes are located on the _____
chromosome. Very few genes are located on the ____ chromosome.
14.An example of a disease caused
chromosome is:

by an allele carried on the X-

15.Two examples of polygenic diseases are

and
16.One of the most common human chromosome disorders, which
involves having an extra 21st chromosome, is called
17.Turners Syndrome in
females is a result of
______________________________________.
18.An extra X chromosome in males results in
__________________________, the symptoms of which are usually
treatable through ________________ and __________________.
19.Most of us carry _________ lethal recessive genes.

Laboratory 8: Mitosis and Meiosis 15

LAB SUMMARY
Please include the following in your lab summary.
1.

Descriptive title

2.

Introduction describing purpose and objectives of this lab activity. Also very
briefly describe the general approach taken to achieve these objectives.

3.

For the first experiment (Mitosis in the Onion Root Tip) include the
following:
Draw labeled diagrams that illustrate the stages of mitosis in the onion root
tip.
Table that lists the number of cells observed in different stages of mitosis.
Assuming that the length of time spent in each stage is proportional to the
number of cells in that stage calculate the duration of each stage of mitosis
(assume mitosis takes 30 minutes).
A paragraph describing the trends observed. Also include comment whether
or not the results made sense and why. Comment on any sources of error.

4.

For the second experiment (Meiosis in the Lilium Anther) draw labeled
diagrams that illustrate the stages of meiosis.

5.

For the third experiment (Giant Chromosomes of Drosophila Larvae) draw a

labeled diagram that illustrates the chromosome.

6.

Conclusion that summarizes the experiments and interprets the significance of

the results.

7.

Attach filled-in (with your answers) pages for QUESTIONS TO

RESEARCH AND ANSWER on pages 13 & 14 of your lab description.