BIOL227

Course introduction &

Introduction to the
compound microscope
A few words on Learning
Outcomes
Learning outcomes in 200-level courses
are mostly restricted at the “knowledge”
and “comprehension” level cognitive
skills.

Courses at the 400-level: Include

“evaluation” and “synthesis”-level cognitive
skills.

Professors/researchers/professionals continue to
hone their synthesis-level skills.

How good you are at the “application” to

“synthesis” level depends on how solid
your foundation is at the knowledge and
Broad learning outcomes for BIOL
227
1. List unique features (at the organismal, cellular and subcellular level) of a particular taxon
2. Identify/label these features (e.g. cell morphology, cell walls, food/contractile
vacuoles, vasculature in plants, etc.)
3. Describe these features and explain why they are important. What advantage do they
confer to the organism? What information does it give you in terms of the organism's
environment and evolutionary history?
4. Recognize the diversity of organisms and their unique features across various levels of
the taxonomic hierarchy.
5. Compare taxa within a taxon (e.g. various classes within a phylum).
 Why do these learning outcomes
matter?
These cognitive skills allow us
• to make better arguments for the role biodiversity plays on our planet.
• to formulate and defend hypotheses about a taxon's evolutionary
history.

• to pick better model organisms for research on human diseases

(i.e. create/develop better research questions)

In short, never underestimate the power of clear and well understood

basic knowledge
The compound light microscope: Learning
Outcomes
1. Describe the parts of the compound microscope (see hand out “How to use a
microscope”)

2. Describe the bending of light as it moves through the lenses (condenser, objective
and ocular lenses) and media (air, water and oil). (In fewer but more technical
terms: Describe the refraction index of light)

3. Explain how an increase in resolution is achieved as we move from scanning to high-

power objective lenses. (In fewer but more technical terms: Explain the Abbé
equation).

4. Practice the Köhler illumination step, oil immersion and dark-field illumination
The refraction index of
light
When light passes from one medium to another, refraction
occurs.

The medium can be air, glass or oil. When light crosses through
a medium with a different density, its speed changes.
Consequently, its path changes. This change in direction is
called refraction.

The refraction index is a measure of how much a medium slows

the velocity of light
• Refractive index of air=1

• Refractive index of glass=1.52 (Source: Willey, Joanne M., Sherwood, Linda., Woolverton, Christopher J., Prescott, Lansing M.
(2008). Prescott, Harley, and Klein's microbiology (7th). McGraw Hill.)
The direction and magnitude of the bend is a function of
• Refractive index of immersion oil=1.52
the refractive indices at the interface of the two media
 Lenses – a microscope’s
jewels.
Three lenses:

1. Condenser lens

2. Objective lens

3. Ocular lens

The objective and ocular lenses magnify the

image. Hence why we call it the
“compound” microscope.
The condenser lens (a collection of
https://upload.wikimedia.org/wikipedia/commons/2/27/
(Source: Willey, Joanne M., Sherwood, Linda., Woolverton, Condenser_diagram.svg
Christopher J., Prescott, Lansing M. (2008). Prescott, Harley,
and Klein's microbiology (7th). McGraw Hill.)

prisms) converges rays of light to a focal

point (F).
Explaining the Abbé
equation
Resolution is the ability of a lens to separate or distinguish
between small objects that are close together.

Ernst Abbé (1870s) a physicist worked out the resolution of

lenses
mathematically. Abbé
equation:
d = 0.5 or, d = 0.5 
n NA
sin
Where,

d= The minimal distance between two points that can be resolved. The
resolving power of the objective lens.

= the wavelength of light

n= refractive index of the medium in which the objective lens is working.

Sin  = 1/2 the angle of the cone of light entering the objective

lens Numerical aperture (N.A) = n sin

(Source: Willey, Joanne M., Sherwood, Linda., Woolverton, Christopher J., Prescott, Lansing M. (2008). Prescott, Harley, and Klein's microbiology (7th). McGraw Hill.)
From Abbé to the importance of doing the Köhler illumination
step To maximize the resolution of a microscope, the Kohler illumination protocol is performed. The
Köhler illumination ensures that the cone of light emerging from the condenser lens is
centered in your field of view. This enables a more uniform illumination across the specimen.
Consequently, both the contrast and resolution improve. The opening and closing of the
aperture iris diaphragm controls the size of the cone of light emerging from the condenser
lens.

Inverted cone of light entering the objective lens

Cone of light emerging from condenser lens and striking the specimen on
slide

Aperture iris diaphragm (or condenser diaphragm)

Circle of light crossing up and through the aperture iris diaphragm

Controlling for
glarea dark-field stop is fitted to the aperture iris diaphragm
When
we are observing the specimen under dark-field.
Dark-field microscopy is used for viewing live specimens.
Only light that is reflected or refracted by the specimen enters
the objective lens. The background appears dark, hence why it
is called dark-field.
Dark-field microscopy improves contrast of live specimens and
also prevents them from overheating.

Dark-field stop