UNIVERSITY OF BELIZE

FACULTY OF SCIENCE & TECHNOLOGY

DEPARTMENT OF SCIENCE
SEMESTER 1, 22-23

CYTOLOGY LAB (BIOL 1151L)

LABORATORY 2- MICROSCOPE CALCULATIONS DATES: 22/09/22, 23/09/22
LAB MANUAL

Calculation of total magnification, the diameter of the field of view, estimation of object size, drawing
magnification, drawing a scale bar

Objectives
 To calculate the total magnification of an object viewed through the ocular lens
 To calculate the diameter of the field of view at scanning, low and high power objectives
 To draw a biological drawing
 To estimate the size of an object
 To estimate the magnification of the drawing
 To draw a scale bar
 To calculate the magnification of the drawing and estimate the size of the object using a scale bar

Procedure
View the following videos from the links given below before coming to the lab

To learn the rules of biological drawing, conversion of units of cell measurements, and drawing a scale bar,
view the video ‘Drawing Scaled Diagrams of Cells’ from the link
https://www.youtube.com/watch?v=4c5539urfyM

To learn how to calculate total magnification and know the relationship between Magnification, Actual size,
and Image size, view the video ‘IGCSE. 2.3 Calculating magnification of drawings’
https://www.youtube.com/watch?v=DPwhJ8FJ9LU&t=61s

To learn how to calculate the field of view, view the video ‘Microscope measurements’
https://www.youtube.com/watch?v=An5Aq3GqRmM

Size of cells
Cells are small for practical reasons and the units of measurement used in cytology are micrometres (µm) and
nanometres (nm).

Table 1: Metric units commonly used in Cytology

The basic unit of measurement of length in the metric system is the meter.
There are 1000 millimeters (mm) in one meter.
There are 1000 micrometers (µm or microns) in one millimeter.
There are 1000 nanometers (nm) in one micrometer.
How to convert between centimetres (cm), millimetres (mm), micrometres (µm) and nanometres (nm)

1. How to calculate the total magnification of a microscope?

When you are seeing an object through the eyepieces of a compound microscope, the image that you are
actually seeing is being magnified twice: by the eyepieces and the objective lens you are using to view an
object.

Therefore, you need to know how to calculate the total magnification of an object seen when using the
compound microscope.

The total magnification of an object observed through the eyepieces (ocular lens) is calculated by multiplying
the ocular lens magnification, which is usually 10X times the magnification of the objective lens being
used (either: 4X, 10X, 20X, 40X, 100X, etc.). “X” is placed after obtaining the total magnification.

Example:
The formulae for Total Magnification (T.M) = Ocular lens magnification x objective lens magnification
Ocular lens magnification = 10X Objective lens magnification being used = 4X
What is the total magnification?
Calculation: T.M. = (10X) x (4X) = 40X

2. How to calculate the diameter of the Field of View (dFOV) of a Microscope?

The microscope field of view (FOV) is the maximum area visible when looking through the microscope
eyepiece.

To estimate the field of view under the scanning objective, you have to place a transparent ruler on the
microscope stage and count the number of divisions on the ruler spanning the FOV.
Looking at the diagram below, the diameter of the field of view under scanning objective lens (4X) with a Total
Magnification of 40X is approximately 4mm.

Calculating the diameter of the field of view (dFOV) for low power objective (100x) or high power
objective (400x)
There is a relationship between the diameter of field of view of a specific objective lens with the diameter of the
field of view of another objective lens.

𝑑𝐹𝑂𝑉 #1 × 𝑚𝑎𝑔𝑛𝑖𝑓𝑖𝑐𝑎𝑡𝑖𝑜𝑛 #1 = 𝑑𝐹𝑂𝑉 #2 × 𝑚𝑎𝑔𝑛𝑖𝑓𝑖𝑐𝑎𝑡𝑖𝑜𝑛 #2

i.e

dFOV of scanning X magnification of scanning lens =dFOV of low power objective X magnification of low power objective

similarly

dFOV of scanning X magnification of scanning lens =dFOV of high power objective X magnification of high power objective

Since we have already estimated the diameter of the field of view for the scanning lens and it shows to be 4mm,
then we can use this information to calculate the diameter of field of view of either the low power or high power
objective lenses.
For the low power objective (10x)
dFOV of scanning x magnification of scanning lens = dFOV of high power objective x magnification of high power objective

(4mm) x (4x) = (dFOV low power) x (10x)

Transpose the 10x to the left and the unknown, which the dFOV low power, remains to the right

(4mm) x (4x) = (dFOV low power)

(10x)

1.6 mm = dFOV low power

Convert the 1.6 mm to um, i.e multiply by 1000. Will give 1600 um.
Therefore, dFOV of low power = 1600 um.

3. How to estimate the size of an object?

Knowing the dFOV allows you to find the size of the object you are observing. To estimate the object size, you
will first need to estimate how many times the object fits across the dFOV. Looking at the example below, you
can estimate that the object fits across our dFOV 2.25 times at high power.
Once you have determined the dFOV, you can estimate the size of the object you are viewing under the
microscope. To find the object size you will have to estimate how many times the object fits across the dFOV.

𝑑𝐹𝑂𝑉 (𝑖𝑛 µ𝑚)

𝑂𝑏𝑗𝑒𝑐𝑡 𝑠𝑖𝑧𝑒 =
𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑡𝑖𝑚𝑒𝑠 𝑜𝑏𝑗𝑒𝑐𝑡 𝑓𝑖𝑡𝑠 𝑎𝑐𝑟𝑜𝑠𝑠 𝑡ℎ 𝑒 𝑓𝑖𝑒𝑙𝑑 𝑜𝑓 𝑣𝑖𝑒𝑤

To estimate the average cell length and breadth

4. How to calculate the drawing magnification?

If you are asked to draw a diagram of the object you are observing, you can calculate the drawing
magnification. This is the number of times your drawing has been enlarged relative to the true size of the
object. To calculate the drawing magnification, use the following equation:

𝑑𝑟𝑎𝑤𝑖𝑛𝑔 𝑠𝑖𝑧𝑒
𝐷𝑟𝑎𝑤𝑖𝑛𝑔 𝑚𝑎𝑔𝑛𝑖𝑓𝑖𝑐𝑎𝑡𝑖𝑜𝑛 =
𝑜𝑏𝑗𝑒𝑐𝑡 𝑠𝑖𝑧𝑒

Note: this equation requires the drawing size and object size to have the same units of measurement.

Example:
If a student draws an image of the animal cell that he/she observed under the microscope and the animal cell
was 200 µm in size (this is the estimated size of the object) under the microscope and his/her drawing on his/her
lab notebook is 3 cm (3000 µm) in size (this is the final size of the object), how many times has he/she enlarged
the object?
Answer: x = 3000 µm / 200 µm = 15. Therefore, after drawing the image in his/her notebook, the magnification
of the image is x15.

5. How to draw a scale bar for a biological drawing?

A scaled diagram is a scientific drawing of a specimen (cell in this case) that has an appropriate scale to
indicate its actual size. A scale bar is a straight line that represents the relationship between the space on your
page and the actual space occupied by the specimen. Draw a 1cm line below the drawing.

Scale= Estimated size (µm)

Drawing size (mm)

The bacterial cell shown below has a scale bar of 10mm (1cm) = 1 µm
Using the scale bar, you can calculate the magnification of the image or drawing and estimate the size of
the specimen.

6. How to calculate the magnification (x) of an image or drawing using a scale bar?

1. Measure the scale bar image (beside drawing) in mm.

2. Convert to µm (multiply by 1000).
3. Magnification = scale bar image divided by actual scale bar length (written on the scale bar).

From the example below, the actual length of the scale bar is 100µm

100µm
 7. How to calculate the size of a specimen using its scale bar?

By cross multiplication (using scale bar)

If 32 mm in picture = 100µm of actual size
Therefore, 83mm of object in picture = actual size of beetle unknow (what we want to calculate)

So, by cross-multiplication

Actual size of beetle unknow = 83mm of the object in picture x 100µm of actual size
32 mm in picture

= 259.375 µm