Observing mitosis *warming is done after staining to intensify the stain
To see mitosis in action you need to look at living cells.
Garlic bulbs grow roots that have actively diving cells in
their tips. Each cell has only eight chromosomes so it is
relatively easy to see the chromosomes once they have
condensed.
In order to see the chromosomes inside the cells, the cells
must be separated and spread out in to layer that is ideally
just one cell thick. Plant cells are glued together by the
middle lamella of pectin proteins. Hydrochloric acids will
break down the pectins that hold the cell together.
1. Cut 1-2cm of the tip from a growing root, e.g. Garlic
2. Place the root tip on a watch glass and add one drop of
warm hydrochloric acid and leave it for 5 minutes. *
3. Add 10 drops of toluidine blue (or acetic orcein)
4. Warm the watch glass on a hot plate for 5 minutes. *
5. Cut about 4-5mm from the growing tip, keep the
rounded tip and discard the rest. *
6. Place the root tip on a microscope slide and using a
mounted needles break open the root (maceration).
7. Cover with a coverslip and blot well by placing the slide
between the fold of a filter paper.
8. Press gently on the coverslip using your thumb to spread
out the root tip.
9. Observe under the microscope and look for cells
undergoing mitosis.
Notes:
*hydrochloric acid is used to break down the pectin in the
middle lamella so that the cells will slide past each other
when squashing
*rounded tip contains the actively diving cell and so does
the white tips than the brown ones.
Mitotic index = number of cells showing stages of
mitosis
total number of cells in the field of
view
Explain how you ensure, or could ensure, that the
results are reliable and valid.
Reliable results are produced when repeated measurements
give similar values. Students need to explain that they have,
or could in the future, take repeated measurements and
calculate the mean.
Valid results measure what they are supposed to do. Validity
can be ensured by carefully using an eye piece graticule
which has been calibrated correctly using a stage
micrometer.
Suggest a reason for having very few dividing cells in
your root tip.
The actively diving apical meristem of the root was not
being viewed. The root used no longer had dividing cells.
Safety precautions
Wear eye protection
Toluidine blue will stain skin and clothes
Orcein ethanoic stain is corrosive, cause severe
burns, has an irritating vapour and stains
If contacted with the skin immediately wash
thoroughly for 10 minutes