Name: Date:

Section:

Mitosis in Onion Root Tips

Laboratory Report 6

Objective: Determine the time spent in different phases of the life cycle

Directions: The cell's life cycle is typically divided into 5 major phases. Draw the phases and describe the
major events that occur during each phase.

Pre Lab Activity

I. Draw and label the onion root tip

II. Label the following micrographs accordingly, showing each stage of mitosis. (Note: The micrographs
are not arranged in the PMAT order)
Materials:

Chemical resistant gloves

Hydrochloric acid solution, HCl, 1 M

Forceps

Methylene blue stain solution 1%

Microscope slides, glass

Onions, bulbs or green

Paper towels

Water, deionized and tap

Pencil with eraser

Compound microscope

Pipets, 3

Cover slips

Ruler

Cup

Scalpel

Safety Precautions

Hydrochloric acid solution is toxic by ingestion or inhalation and corrosive to skin and eyes. Methylene
blue stain is a permanent stain on many objects. The scalpel is a sharp object—use care when cutting the
root tips with the scalpel. Wear chemical splash goggles, chemical resistant gloves, and a
chemical-resistant apron throughout this lab. Wash hands thoroughly with soap and water before
leaving the laboratory. Follow all normal laboratory safety guidelines.

Procedure

1. Cut three roots from an actively growing plant using a scalpel. Caution: The scalpel is extremely sharp.
2. Trim the tip of each root to 1 cm; use only the tapered end of the root tip.

3. Use forceps to place 2–3 root tips (use only the 1-cm tips) on a glass microscope slide.

4. Using a clean, graduated pipet, add 2–3 drops of 1 M hydrochloric acid to cover the root tips on the
microscope slide. Note: Hydrochloric acid is corrosive to skin and eyes.

5. Allow the root tips to soak in the acid for 5 minutes.

6. After 5 minutes, use a paper towel and carefully blot away excess hydrochloric acid from the slide.
Caution: Avoid contact of the acid with skin.

7. Using a clean, graduated pipet, add 2–3 drops of deionized water to the root tips.

8. Use a paper towel to blot away excess water.

9. Repeat steps 7 and 8.

10. Using a clean, graduated pipet, add 2–3 drops of methylene blue stain to the root tip. Note:
Methylene blue stain is a permanent stain.

11. Allow the root tips to soak in the stain for 3 minutes.

12. Use a paper towel to blot away excess methylene blue stain.

13. Add 1 drop of deionized water to the root tips.

14. Use forceps to move one root tip to a clean microscope slide.

15. Place a cover slip on the root tissue. Using the eraser end of a pencil, gently apply pressure on the
cover slip to squash the root tissue. Apply an even downward pressure on the root tips and cover slip but
not so hard as to break the cover slip. Do not twist or grind the cover slip.

16. Using low magnification on the microscope, focus on the root cells. Switch to medium power or high
power as necessary to easily visualize the inside of the onion root cells.

17. Study all of the squashed tissue to locate cells in each stage of the cell cycle. Note: All stages of
mitosis may not be present within a single field of view.

18. Repeat steps 14–17 using the remaining two root tips.
Intro
Ever wonder how we grow physically? One key factor is the cell cycle! Here's a cool fact: cells
divide much faster in early embryos, about every 30 minutes, compared to the typical 24-hour cycle in
adults. The cell cycle is a series of events that lead to cell division and duplication, essential for growth
and repair. It consists of the interphase (G1, S, and G2 phases) and the mitotic phase (mitosis and
cytokinesis). During interphase, the cell grows, replicates DNA, and prepares for division. Mitosis is the
process where the cell's nucleus divides, followed by cytokinesis, which splits the cell into two. This cycle
is a tightly regulated process to ensure accurate DNA replication and distribution (Earnshaw et al., 2022).

In this lab activity, the scientists were tasked with using the onion root tips they grow to observe
how the cell cycle works, but due to several setbacks, they were ordered to use a prepared sample of
onion root cells instead. After observing, another task was given to the scientists which was describing
the cell cycle phases using feature tables.
Analysis:

Features Interphase Karyokinesis Cytokinesis

Cell morphology The cell is larger Prophase The cell fully
with a distinct The cell maintains its shape, but the nucleus splits into two
nucleus begins to break down daughter cells
Metaphase
The cell appears slightly elongated as
chromosomes align
Anaphase
The cell elongates further as chromosomes are
pulled apart
Telophase
The cell begins to pinch in the middle (cleavage
furrow forms)
Nuclear The nuclear Prophase Two separate
morphology envelope is intact, The nuclear envelope dissolves, and nucleoli nuclei are fully
and nucleoli are disappear visible
visible Metaphase
The nuclear envelope has fully disappeared
Anaphase
No nuclear envelope present
Telophase
Two distinct nuclei begin to form, nucleoli
reappear
Chromosomes Chromosomes are Prophase Chromosomes
not distinct; they Chromosomes condense and become visible as have fully
exist as chromatin, individual structures. decondensed
in a diffused state. Metaphase into chromatin
Chromosomes are aligned at the cell's within each
equatorial plate. nucleus.
Anaphase
Sister chromatids are separated and pulled
toward opposite poles.
Telophase
Chromosomes begin to decondense into
chromatin.
The scientists observed the prepared sample and described the following features: cell
morphology, nuclear morphology, and chromosomes. Initially, for cell morphology, they struggled to
identify the interphase but eventually recognized it upon seeing a distinct structure they speculated to
be the nucleus. For karyokinesis, which includes the phases of mitosis (prophase, metaphase, anaphase,
and telophase), they observed a broken structure under the microscope, which they identified as a cell in
prophase. Elongated, fishbone-like structures were interpreted as cells in metaphase, while a crescent
moon-like structure, more elongated, was considered to represent a cell in anaphase. Finally, two distinct
black structures with a visible line in between, which they believed to be a cleavage furrow, indicated a
cell in telophase. During cytokinesis, the scientists were unable to observe cells fully splitting into two
daughter cells, as the microscope only showed individual cells, implying the process had already been
completed.

In terms of nuclear morphology, in the identified interphase cell, the nuclear envelope might
appear slightly blurred under the microscope but is still recognizable as intact, with the nucleoli visible.
During karyokinesis, starting with prophase, the nuclear envelope is no longer recognizable, and the
nucleoli seem to be rupturing. In metaphase and anaphase, the nuclear envelope completely disappears,
and the nucleoli are no longer visible in either phase. In telophase, two distinct black structures were
observed, which the scientists speculated to be newly forming nuclei. For cytokinesis, two fully
separated and visible nuclei are expected, but this was not observed, as the cells in the sample appeared
as individual, fully-formed cells.

Regarding the features of chromosomes, during interphase, chromosomes are not visible within
the nucleus, which appears as a black structure under the microscope. The scientists speculated that this
is because chromosomes in interphase exist as chromatin, in a diffused state, making them hard to
distinguish. In karyokinesis, starting with prophase, chromosomes become visible as individual, more
condensed structures. In metaphase, the chromosomes can be seen aligned along the equatorial plate,
which, based on cell morphology, resembles fishbone-like structures. During anaphase, the
chromosomes are pulled apart by crescent-shaped structures, which the scientists identified as spindle
fibers pulling sister chromatids toward opposite poles. In telophase, chromosomes are observed to
decondense back into chromatin as the nuclei begin to reform. During cytokinesis, the chromosomes
were not directly visible, as they were likely inside the newly formed nuclei, which appeared as black
structures. However, the scientists inferred that the chromosomes had fully decondensed into chromatin
inside the nucleus at this stage.

Conclusion:

In this experiment, the scientists studied the cell cycle using prepared onion root cell samples,
successfully identifying the stages of interphase, karyokinesis (prophase, metaphase, anaphase, and
telophase), and cytokinesis. They observed the distinct changes in cell morphology, nuclear structure,
and chromosome organization that occur throughout these stages, such as the breakdown of the nuclear
envelope and chromosome alignment during metaphase. Although there were challenges in recognizing
interphase and observing the completion of cytokinesis, the scientists were able to analyze and
document the key phases. This experiment reinforced the significance of the cell cycle in growth and
development.

Post Lab questions - (GCLASS IPAPASA)

IV. Summarize the major differences between mitosis and meiosis in the table below:

Mitosis Meiosis
Crossing over
When chromosomes split
Number of divisions
Number of cells resulting
Number of chromosomes in
daughter cells
 Genetic similarity of daughter
cells to parent cells

V. Reviewing Your Knowledge

(1.) Define the following terms:

1.centromere:

2. centriole:

3. centrosome:

4. kinetochore:

5. sister chromatid:

6. cell plate:

7. cleavage furrow:

8. diploid/haploid:

(2.) What role would mitosis play in the body of an adult animal?

(3.) What advantage does the process of crossing over bring to reproduction?

(4.) Why would the method of cytokinesis in animal cells not work in plant cells?

(5.) How is Prophase I in Meiosis different from Prophase in Mitosis. Explain in detail.

(6.) What is the genetic significance of crossing over?

VI. Explain synapsis and crossing over. Sketch a diagram to represent both.
VII. Complete and answer the table below.

Mitosis Meiosis
What is the purpose of this
process?

What is the outcome of this

process?

Which organisms perform this

process?

How long does this process

take?

What is an example of a disease

caused by an error in this
process?