NATIONAL UNIVERSITY OF SCIENCE AND TECHNOLOGY
FUCULTY OF AGRICULTURE
NAME : TINOTENDAISHE B
SURNAME: NYACHIYA
STUDENT NUMBER: N02537309A
PROGRAME: AGRICULTURAL GENETICS AND CELL TECHNOLOGY
COURSE: CELL BIOLOGY
LECTURER: MR MUSUSA
TITLE: TYPES OF MICROSCOPES
1 Compound Microscope
Compound Microscope is a type of microscope that used visible light for illumination and
multiple lenses system for magnification of specimen. Generally, it consists of two lenses;
objective lens and ocular lens. It can magnify images up to 1000X. Its magnifying power is equal
to the product of magnifying power of the objective lens in use and the ocular lens.
Compound Microscope Working Principle
When light is focused through a condenser on a specimen placed on stage, the light transmitted
by the specimen is picked by the objective lens. A magnified image is formed at the body tube.
This is called the primary image. The light bends in the body tube and passes through the ocular
lens. When passing through the ocular lens, the image is magnified for the second time. This is
called the secondary image. Finally, a highly double magnified image is formed at a distance of
distinct vision.
Compound Microscope preparation
- Fixation: Tissue is preserved using chemicals like formalin to prevent decay.
- Dehydration: Water is removed using alcohol solutions.
- Embedding: Specimen is embedded in paraffin wax to support slicing.
- Sectioning: Thin slices (5–10 µm) are cut using a microtome.
- Mounting: Sections are placed on glass slides.
- Staining: Dyes like hematoxylin and eosin are used to highlight structures
Mostly used in:
- Biology classrooms – for teaching cell structure and basic histology.
- Clinical labs – to examine blood smears, bacteria, and tissue samples.
- Pathology labs – for diagnosing diseases through stained tissue sections.
- Microbiology – identifying microorganisms like bacteria and protozoa.
Pros
-Can achieve magnifications typically ranging from 40x to 1000x, allowing detailed observation
of small specimens.
-Suitable for a wide range of applications in biology, medicine, and education, including
histology, microbiology, and pathology.
-Can be used to observe live specimens, making it valuable for studying biological processes in
real time.
-Generally more affordable than advanced microscopes like electron microscopes, making it
accessible for many laboratories and educational institutions.
-User-friendly design makes it easy for beginners to operate with minimal training.
Cons
- The maximum resolution is around 200 nanometers, which may not be sufficient for visualizing
very small structures (e.g., viruses).
- Some specimens require staining or other preparation methods, which can alter their natural
state and potentially affect observations.
-Limited depth of field can make it challenging to focus on thicker specimens or structures at
varying depths.
-Adequate lighting is essential; poor lighting can affect visibility and image quality.
-Generally only suitable for thin specimens; thicker samples may not be adequately illuminated
or resolved.
-Can’t produce image of objects smaller than wavelength of visible light (0.4 μm)
Electron microscopes
Electron Microscope is a microscope that uses accelerated electron, beams instead of light rays,
to illuminate the specimen and get the highly magnified image. In this microscope, glass lenses
are replaced by electromagnets. Due to the very short wavelength of electrons, this microscope
produces a very high-resolution image with magnification up to 10,000,000X. Very high-quality
images with very high contrast, revealing detailed structures are produced. Specimen up to 0.2
nm can be clearly viewed using an electron microscope.
Electron Microscope Principle
An electron microscope uses accelerated electrons with a wavelength of about 100,000 times
shorter than visible light to illuminate specimens and produce images. The electron gun, usually
a heated tungsten or field emission filament, is used to generate a stream of high voltage (100 –
1000 kV) electrons. These electrons are accelerated using an anode plate in a vacuum system and
focused on the specimen using aperture and electromagnetic lenses.
The electron beam passes through the specimen and interacts with sample components. Upon
striking the specimen, the electrons are scattered. The degree of scattering depends on the
refractive index or thickness of the specimen.
The scattered electrons from the sample are collected and passed through objective and ocular
electromagnetic lenses. These scattered beams are detected and transformed into highly
magnified images by the magnetic lenses.
Preparation Steps:
- Fixation: Glutaraldehyde followed by osmium tetroxide to preserve and contrast cellular
structures.
- Dehydration: Alcohol or acetone series to remove water.
- Embedding: Specimen is embedded in epoxy resin.
- Sectioning: Ultra-thin slices (~70 nm) are cut using an ultramicrotome.
- Mounting: Sections are placed on copper grids.
- Staining: Heavy metals like uranyl acetate and lead citrate are used to enhance contrast.
- Final Thinning (for metals):
- Bulk samples are sliced to ~150–300 µm.
- Ground to <100 µm.
- Punched into 3 mm discs.
- Dimpled to 2–10 µm.
- Jet-polished or ion-milled to achieve electron transparency.
Mostly used in:
- Cell biology and virology – visualizing organelles, viruses, and macromolecules.
- Cancer research – examining cellular ultrastructure and abnormalities.
- Materials engineering – studying atomic arrangements and defects.
- Pharmaceuticals – analyzing drug delivery systems and nanoparticles.
Pros of Electron Microscopes
- Can magnify up to 2 million times, revealing atomic-level detail.
- Far superior to light microscopes due to shorter electron wavelengths.
- Produces highly detailed images of surface structures (SEM) and internal ultrastructure (TEM).
- Ideal for studying viruses, organelles, nanomaterials, and crystal lattices.
- Used in biology, medicine, materials science, forensics, engineering, and semiconductor
inspection.
- Enables breakthroughs in nanotechnology, drug development, and metallurgy.
4. 3D Imaging (SEM)
- Scanning Electron Microscopes provide three-dimensional views of specimen surfaces.
Cons of Electron Microscope
- Extremely expensive to purchase and maintain.
- Requires specialized infrastructure and shielding from vibrations and magnetic fields.
- Specimens must be dehydrated, fixed, embedded, sectioned, and stained—especially for TEM.
- Preparation can introduce artifacts that distort the image
- Cannot observe living specimens due to vacuum environment and electron beam damage.
- Large, bulky equipment that’s sensitive to environmental conditions.
- Needs stable voltage, cooling systems, and trained personnel.
