Department of Biology and Microbiology, “Molecular and Cell Biology”

Group
Name 2
Subhaan
Surname Date
Chaudhry 23.02.2023

PROTOCOL No.1.

MICROSCOPY OF NUCLEUS AND HUMAN METAPHASE CHROMOSOMES

Task I: Look at the light microscopy slides under the large objective (blue, x40) and take photographs of
them (when possible, use smartphone camera):
a) Frog erythrocytes;
b) Human erythrocytes;
c) After the practical class cut and paste the images (from your camera or the e-studies folder “Practical class
No 1_materials”) into the protocol.
d) Answer the questions below.

Frog erythrocytes, magnification 400x Human erythrocytes, magnification 400x

Question Answer
1.1.Compare frog and human erythrocytes by size and The frog erythrocytes Are larger than human
presence or absence of the nucleus erythrocytes and contain a nucleus whereas human
erythrocytes do not.

1.2. How does the shape of the erythrocytes influence The erythrocytes have a bioconcave disk shape
their functions? which gives them a large surface area for oxygen
absorption. This shape also provides flexibility and
allows erythrocytes to squeeze easily through blood
vessels.

1.3. Why do human and frog erythrocytes differ from
each other?
Due to environmental differences frogs and humans
will have different affinities for oxygen and
therefore have different erythrocyte adaptations.
Differences in metabolic rate and demand for
oxygen also contributes to differing adaptations.

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 Department of Biology and Microbiology, “Molecular and Cell Biology”

Task II: Analysis of metaphase plate by using oil immersion objective:

a) Observe the sample in small objective (yellow, x10);
b) Find a preferable metaphase spread in large objective (blue, x40);
c) Apply immersion oil and observe the chromosomes under immersion objective (white, x100). Take pictures of the
chromosomes
After the practical class cut and paste into the protocol images (from your camera or the e-studies folder “Practical class
No 1_materials”) that show a metacentric, submetacentric and acrocentric chromosome. Using the Snipping tool, Paint
or similar software, cut the respective chromosomes from the provided image and past them in the corresponding cell
in the table below. Label the centromere, long arm (q) and short arm (p) for each chromosome.

Metacentric chromosome Label

1 - centromere
2 – long arm (q)
3 – short arm (p)

Submetacentric chromosome Label

1 - centromere
2 – long arm (q)
3 – short arm (p)

Acrocentric chromosome Label

1 - centromere
2 – long arm (q)
3 – short arm (p)

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 Department of Biology and Microbiology, “Molecular and Cell Biology”

Task III: Based on your knowledge, answer the questions in the table below.
Question Answer
3.1. Why do we analyse human chromosomes
during metaphase, but not e.g. during interphase?
During metaphase, there is the highest level of
chromosome condensation and so is most
distinguishable. However during interphase there is
the least condensation and so is not as visible.

3.2. Can you make conclusions about the changes

in number or structure of certain genes by analysis
of microscopy slide of metaphase chromosomes?
Motivate your answer.
Analysis of metaphase chromosomes can tell us the
size, structure and the number of chromosomes.
However, conclusions cannot be made about the
number or structure of certain genes because they
are located on chromosomes and are too small to
see with a light microscope and require an electron
microscope to be seen and analysed.

Task IV: Female is at 15th week of a pregnancy. During first trimester screening (using biochemical and
ultrasonography data), the doctor gets suspicious about possible chromosomal pathology in the foetus. The
doctor does amniocentesis (taking the sample of amniotic fluid) and the sample is referred to the cytogenetic
laboratory. After cultivation of the cells and chromosome staining, you can see following image in the
microscope (see below).
Question Answer
4.1. Write the karyotype of foetus. 45, XO

4.2. Does the result confirm suspected diagnosis of Yes

chromosomal pathology?
4.3. To which type of chromosomes (depending on Chromosome X – Submetacentric

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 Department of Biology and Microbiology, “Molecular and Cell Biology”

the localization of the centromere) does sex

chromosome X and sex chromosome Y belong Chromosome Y – Acrocentric
to?

Karyotype used with permission of cytogeneticists from E. Gulbis laboratory.