Required Practical 2

Mitotic Index
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Introduction
Method
Risk Assessment
Results
Analysis
Should be typed
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Competencies Tested:
1a, 2b, 3a, 4a and 5a
1a. Correctly follows written instructions to carry out experimental techniques or
procedures.
2b. Carries out techniques or procedures methodically, in sequence and in
combination, identifying practical issues and making adjustments when necessary.
3a. Identifies hazards and assesses risks associated with these hazards, making safety
adjustments as necessary, when carrying out experimental techniques and procedures
in the lab or field.
4a. Makes accurate observations relevant to the experimental or investigative
procedure.
5a. Uses appropriate software and/or tools to process data, carry out research and
report findings.
 Write- up
3a. Identifies hazards and assesses risks associated with these hazards,
making safety adjustments as necessary, when carrying out experimental
techniques and procedures in the lab or field.

Risk Assessment Table

Risk Why it is a risk What I will do to
overcome the risk
 Write- up
4a. Makes accurate observations relevant to the experimental or
investigative procedure.
- Picture with stages circled and identified
- Mitotic Index Calculated
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Competencies Tested:
5a. Uses appropriate software and/or tools to process data, carry
out research and report findings.

- Picture
- Using excel to calculate mitotic index
- Research using Google Scholar (include article and date
accessed)
Methodology
1. Heat 1 mol dm-3 HCl at 60°C in a water bath.
2. Cut a small sample of the root tip using a scalpel.
3. Transfer root tip to HCl and incubate for 5 minutes.
4. Remove from HCl and wash sample in cold distilled water and remove the very tip using a scalpel.
5. Place tip on a microscope slide and add a few drops of stain (e.g. toluidine blue O). This makes the
chromosomes visible and will therefore show which cells are undergoing mitosis.
6. Lower the cover slip down carefully onto the slide. Make sure there are no air bubbles in the slide
which may distort the image. Squash the cover slip down using paper (to protect your hands from
dye) to make a thin layer of cells.
7. Place under a microscope and set the objective lens on the lowest magnification.
8. Use the coarse adjustment knob to move the lens down to just above the slide.
9. Use the fine adjustment knob to carefully re-adjust the focus until the image is clear (you can use a
higher magnification if needed).
10. To calculate mitotic index, cells undergoing mitosis must be counted (cells with chromosomes
visible), as well as the total number of cells.
 Write- up
1. Why did you soak your root tips in Hydrochloric acid for 15mins?
2. Why did you use only the final 1mm of the root tip?
3. Why did you use a stain?
4. How did you press firmly on the slide? Why did you do this?
5. How do you calculate mitotic index?
6. What are the limitations of the method?