Observing Mitosis in Onion Root Tips

Purpose: To observe the stages of mitosis in the cells of actively growing onion root tips.

Materials needed for each group:

- Onion root tips - safety goggles - metric ruler - forceps -

microscope
- toluidine blue stain - probe - paper towels - apron - scalpel

- 2 glass slides with plastic coverslips - stopwatch or clock with a second hand

- 3 small beakers (50 or 100 ml) - ethanol/acetic acid fixative (75ml /

25ml ratio)

- 5M Hydrochloric acid (30 ml H2O / 20 ml concentrated HCl) - sharp minds and a focused demeanor

Procedure:

1.) Carelessness, rowdy behavior, or being unable to pay attention to what’s going on around you can get
someone hurt in this lab. Any of these behaviors are unacceptable, and will result in immediate
removal from the lab area. Please get yourself in the correct frame of mind now!

2.) Put on your safety goggles and lab apron. The fixative solution and hydrochloric acid we will be
using during this lab are very corrosive, which means that goggles are required at all times. If you spill
any material, please let me know immediately. You are required to wash your hands when finishing
the experiment.

3.) Make sure that you have 3 beakers, and that they are labeled fixative, hydrochloric acid, and
water.

4.) Bring the beakers up front and obtain 10 ml of each substance. Your teacher will pour these materials
for you.

5.) Using the forceps, place 2 onion root tips into your hydrochloric acid beaker, and let them sit for 4
minutes.

6.) After 4 minutes, remove the tips from the HCl with the forceps and place them in the fixative solution
for 4 minutes.

7.) After 4 minutes, remove the tips from the fixative and place each one on a clean glass slide. Holding
the root tip with the forceps, use the scalpel to cut off about 1mm from each root tip. Use the ruler to
measure this length. CAUTION: the scalpel is extremely sharp, so make sure your finger is
clear when you make the cut. Discard the rest of the root tip, leaving the 1mm piece on your slide.

8.) Place the glass slide on a paper towel, and add 2 drops of toluidine blue stain to the root tip. Allow the
stain to sit for 2 minutes, and then gently use a paper towel to soak up the excess stain surrounding
the root tip. CAUTION: toluidine blue is a stain and will not come out of your clothes – be
careful!

9.) Place 2 drops of water on the root tip and gently cover with a cover slip.

10.) Place a paper towel over the slide, and use the eraser end of your pencil to gently press down on the
cover slip. This will squash the root tip, making the cells more easily viewable. CAUTION: DO NOT
push so hard as to break the cover slip or slide!

11.) Using a microscope, observe the onion root tip cells (remember your microscope skills from 1st
semester!). Look for cells undergoing the different stages of mitosis under med or high power. Draw
 labeled diagrams of these cells on the back of this paper, and call your teacher over to observe these
cells, and to sign your paper.

12.) Clean and dry your microscope slides, cover your microscope and put it away in the cabinet, organize
your lab materials, and wipe down your lab station with a paper towel. Wash your hands using soap
and water for a minimum of 30 seconds!