Name : ___________________________

IBDP Year 1

Practical 1: Light microscopy

Figure 1– Parts of a light microscope

Activity 1: Measuring the field of view (FOV)

 The field of view is the circular area under observation when using a microscope.
 You need to know the diameter of the field of view to calculate the size of the
specimen you are looking at.
 All you need to calculate the field diameter is a slide with a millimetre grid or ruler
mounted on it.

Instructions

1. Mount the mm slide on low power, focus it and measure the field diameter. Remove

the slide from the stage.

Figure 1.2.2b – Millimetre ruler

At 40X magnification, the field diameter of a light microscope normally used in
schools is about 4.0–4.2mm.
 2. The ratio of magnification, M, to diameter, D, is the same at every magnification. Using
the measured diameter from step 1, D1, calculate the diameter of the field of

M1D1 = DX
MX

For example, if the magnification of your ocular lens is 10X and your objective lenses are 4X,
10X and 40X, the field diameters will be 4.2mm, 1.7mm and 0.42mm on low, medium and
high power respectively. Check for yourself!

Activity 2: Preparing a wet mount

Materials

 Plant tissue (e.g. onion)

 Eye dropper
 Iodine
 Glass slide
 Coverslip
 Mounted needle or pencil
 Paper towel

Instructions

1. Peel a layer of plant cells from the sample and lay it carefully on a clean glass
slide.
2. Add a drop of iodine using an eye dropper.
3. Lay one edge of the coverslip down at one end of the specimen. Use a pencil
or mounted needle to gently lower the coverslip over the specimen.

Figure 1.2.2c – Gently lower the coverslip

4. To remove excess water, use a small piece of paper towel at the edge of the
coverslip.
 5. If the slide is too dry, add a drop of water at one end of the coverslip and draw
it through the specimen using the paper towel at the other end of the
coverslip.

Activity 3: Estimating cell size

Once you know the diameter of the field of view, you can calculate the actual size of
the cells you are looking at, using this simple calculation:

Figure 1.2.2d – Onion epidermal cells at 100X magnification

In activity 1, we calculated that the diameter of the FOV at medium power is 1.7mm.
To convert millimetres (mm) to micrometres (μm), multiply by 1000 (1mm = 1000
μm). In Figure 1.2.2d, five cells fit across the diameter of the FOV (lengthwise) and
fifteen cells fit across the diameter (widthwise), using the formula:

Actual cell length = 1700μm/5 cells = 340μm/cell

Each cell is approximately 340μm in length.

Actual cell width = 1700μm/15 cells = 113μm/cell

Each cell is approximately 113μm in width.
Analysis:
Activity 1 : Measuring the field of view ( FOV)
Low power (10x) =

Medium power (40 x) =

High power ( 100 x) =

Activity 3: Estimating cell size

Using ………………….. power

Actual cell length =

Each cell is approximately …………………. in length.

Actual cell width =

Each cell is approximately ………………………… in width.

Conclusion: