Lab Report: Introduction to Electron Microscopy and Its Comparison with
Light Microscopy
Objective:
To understand the limitations of light microscopy in achieving high magnification and to explore
the concept of resolution. Additionally, to introduce the electron microscope (EM) as a solution
for increasing magnification and to compare the different types of electron microscopes.
Part 1: Limitations of Light Microscopy
1. Magnification and Resolution:
o Magnification refers to the ability of a microscope to make objects appear larger.
Light microscopes can magnify objects up to around 1000x their actual size.
However, this is limited due to the wavelength of visible light (400-700 nm).
o Resolution is the ability of a microscope to distinguish between two points that
are close together. Light microscopes are limited by the wavelength of visible
light. As the magnification increases, the resolution decreases beyond a certain
point, meaning objects can appear blurry and indistinguishable.
Exercise:
o Use the light microscope to view a prepared slide at different magnifications (e.g.,
40x, 100x, 400x). As you increase the magnification, note any blurring or loss of
detail.
Discussion:
o Explain that light microscopes are limited in both magnification and resolution
due to the nature of visible light. This means that objects smaller than a certain
size (around 0.2 micrometers) cannot be distinguished with high clarity.
Part 2: Introduction to Electron Microscopy
Electron microscopes overcome the resolution and magnification limitations of light microscopes
by using electrons instead of visible light. Electrons have much shorter wavelengths (less than 1
nm), allowing for much higher resolution.
1. Types of Electron Microscopes:
o Transmission Electron Microscope (TEM): In TEM, electrons pass through a
very thin sample, allowing for detailed internal views of cells and structures at a
much higher magnification (up to 10,000,000x). TEM provides 2D images.
o Scanning Electron Microscope (SEM): SEM uses electrons to scan the surface
of a sample, creating 3D images. It is ideal for observing the surface structure of
objects at a high resolution (up to 1,000,000x).
Part 3: Differences Between Light and Electron Microscopes
Feature Light Microscope Electron Microscope (TEM & SEM)
Source of Imaging
Magnification Up to 1000x TEM: up to 10,000,000x
SEM: up to 1,000,000x
Resolution ~0.2 micrometers TEM: ~0.1 nanometers
SEM: ~1 nanometer
Image Type 2D (flat) images TEM:
SEM:
Specimen(live/dead)
Specimen color
Sample Preparation Simple preparation Complex preparation (samples need to be
(staining, slides) thin for TEM and coated for SEM)
Accessibility Widely available in Expensive and requires specialized labs
most labs
Works in Air column
Part 4: Electron Micrographs Exercise
Since our lab does not have access to electron microscopes, you will be provided with electron
micrographs of samples taken using SEM and TEM.
Instructions:
• Download electron micrographs from a reliable source (e.g., Google).
• SEM Micrograph: Look for images of surface structures.
Bacteria, flagella, Protozoa (foraminiferans), flagella/cilia, Algae (diatoms), Fungi
(spores), Insects (mouth parts/mandibles), Plants (trichomes/plant hair, stomata,
polengrain), Animals (cilia in respiratory tract, RBC),
• TEM Micrograph: Look for images showing the internal structure of cells or viruses.
Nucleus, ER (RER/SER), Golgi apparatus, Lysosomes, peroxisomes, mitochondria,
chloroplast, cytoskeleton, cross section of flagella/cilia (9+2)
Exercise:
1. Paste the SEM and TEM micrographs in the provided space below.
2. Identify and describe the main differences in the images:
o For SEM, focus on the surface features and the 3D nature of the images.
o For TEM, observe the internal structures of the sample, like organelles, viruses,
or other intracellular components.
Discussion:
• Discuss the differences you observe between the two types of electron micrographs.
Highlight how SEM provides surface details in 3D, while TEM reveals internal structure
in 2D.
Part 5: False Coloring in Electron Microscopy
In electron microscopy, false coloring is often applied to micrographs to enhance contrast and to
highlight certain features. This process is similar to the use of staining in light microscopy,
which is done to improve the visibility of cell structures.
Note: While electron microscopy does not use traditional stains, false coloring serves a similar
purpose, emphasizing particular features for easier visualization and analysis.
Exercise:
• Identify any instances of false coloring in the electron micrographs you downloaded.
Discuss how the false colors help highlight specific parts of the sample, such as
organelles or surface features.
