Microscope
A microscope is an instrument used to observe objects that are too small to be
seen with the naked eye. It magnifies tiny structures such as cells,
microorganisms, and tissue components so that they can be studied in detail.
Microscopes are mainly classified into:
1. Light (Optical) Microscope
2. Electron Microscope
o Transmission Electron Microscope (TEM)
o Scanning Electron Microscope (SEM)
1. Light Microscope
Definition
A light microscope is an optical instrument that uses visible light and glass
lenses to magnify small specimens. It is the most commonly used microscope in
laboratories, schools, and hospitals.
It allows scientists to study cells, tissues, microorganisms, and blood
samples.
Principle
The light microscope works on the principle of refraction of light through
glass lenses.
Light passes through the specimen and is bent by the lenses to produce a
magnified image of the specimen.
Magnification occurs through two lenses:
Objective lens
Eyepiece (ocular lens)
Total Magnification =
Eyepiece Magnification × Objective Magnification
Example:
10× eyepiece × 40× objective = 400× magnification
Parts of a Light Microscope
1. Mechanical Parts
Base
Bottom support of the microscope.
Provides stability.
Arm
Connects the base with the body tube.
Used for carrying the microscope.
Stage
Platform where the slide is placed.
Has a hole for light to pass through.
Stage Clips
Hold the slide in position.
Coarse Adjustment Knob
Moves the stage up and down quickly.
Used for rough focusing.
Fine Adjustment Knob
Used for precise focusing.
2. Optical Parts
Eyepiece (Ocular Lens)
Located at the top.
Usually 10× magnification.
Objective Lenses
Different magnification lenses:
4× (Scanning lens)
10× (Low power)
40× (High power)
100× (Oil immersion)
Condenser
Concentrates light onto the specimen.
Iris Diaphragm
Controls the amount of light reaching the specimen.
Mirror or Illuminator
Provides light for viewing.
Working of a Light Microscope
1. Light from the source passes through the condenser.
2. The condenser focuses light on the specimen.
3. Light passes through the specimen.
4. The objective lens magnifies the image.
5. The eyepiece lens further magnifies the image.
6. The observer sees a magnified image.
Magnification and Resolution
Magnification
Ability to enlarge an object.
Maximum magnification of light microscope:
1000× – 1500×
Resolution
Ability to distinguish two points as separate.
Resolution of light microscope:
0.2 µm (200 nm)
Types of Light Microscopes
1. Bright Field Microscope
2. Dark Field Microscope
3. Phase Contrast Microscope
4. Fluorescence Microscope
5. Confocal Microscope
Applications of Light Microscope
Study of cells and tissues
Blood smear examination
Observation of bacteria and parasites
Histopathology studies
Microbiology research
Medical laboratory diagnosis
Advantages of Light Microscope
Simple to use
Less expensive
Specimens can be viewed alive
Minimal sample preparation
Limitations of Light Microscope
Limited magnification
Lower resolution
Cannot see very small structures like viruses
Less detailed images
Electron Microscope
Definition
An electron microscope is a microscope that uses a beam of electrons
instead of light to produce highly magnified images.
Electron microscopes provide much higher magnification and resolution than
light microscopes.
Maximum magnification can reach 1,000,000× or more.
Principle
Electron microscopes work on the principle that electrons have very short
wavelengths.
Short wavelength allows the microscope to achieve very high resolution.
Instead of glass lenses, electromagnetic lenses are used to focus the electron
beam.
Types of Electron Microscopes
1. Transmission Electron Microscope (TEM)
2. Scanning Electron Microscope (SEM)
1. Transmission Electron Microscope (TEM)
Definition
A Transmission Electron Microscope (TEM) is a type of electron microscope
in which electrons pass through an extremely thin specimen to form an image.
It provides detailed images of the internal structure of cells.
Principle
A beam of electrons is transmitted through a thin specimen.
Different parts of the specimen scatter electrons differently.
The transmitted electrons form an image on a screen or detector.
Structure of TEM
Main components include:
Electron Gun
Produces electrons.
Condenser Lens
Focuses electron beam on specimen.
Specimen Holder
Holds the thin specimen.
Objective Lens
Produces the first magnified image.
Projector Lens
Further magnifies the image.
Fluorescent Screen or Camera
Displays the final image.
Working of TEM
1. Electron gun produces electrons.
2. Electrons are focused by condenser lenses.
3. The electron beam passes through the thin specimen.
4. Some electrons are absorbed and others pass through.
5. Objective lens forms an image.
6. Projector lens enlarges the image.
7. Image appears on a fluorescent screen or camera.
Magnification and Resolution
Magnification:
Up to 1,000,000×
Resolution:
0.1–0.2 nm
Applications of TEM
Study of cell organelles
Observation of viruses
Study of protein structures
Nanotechnology research
Material science
Medical research
Advantages of TEM
Very high magnification
Excellent resolution
Detailed internal structure visualization
Limitations of TEM
Expensive instrument
Complex operation
Specimens must be very thin
Cannot observe living specimens
Requires vacuum conditions
2. Scanning Electron Microscope (SEM)
Definition
A Scanning Electron Microscope (SEM) produces images by scanning the
surface of a specimen with a focused electron beam.
It provides three-dimensional images of the specimen surface.
Principle
The electron beam scans the surface of the specimen.
When electrons strike the specimen, secondary electrons are emitted.
These electrons are detected to produce a 3D image.
Structure of SEM
Electron Gun
Produces electron beam.
Electromagnetic Lenses
Focus the beam.
Scanning Coils
Move the beam across the specimen.
Specimen Chamber
Holds the specimen.
Electron Detector
Detects secondary electrons.
Display Screen
Shows the image.
Working of SEM
1. Electron gun produces electron beam.
2. Electromagnetic lenses focus the beam.
3. Scanning coils move the beam across the specimen.
4. Electron beam hits the specimen surface.
5. Secondary electrons are emitted.
6. Detectors collect these electrons.
7. Image is displayed on the screen.
Magnification and Resolution
Magnification:
Up to 100,000×
Resolution:
1–10 nm
Applications of SEM
Surface structure analysis
Study of bacteria and microorganisms
Material science
Forensic science
Semiconductor industry
Medical research
Advantages of SEM
Produces 3D images
High resolution
Large depth of field
Detailed surface structure
Limitations of SEM
Expensive
Sample must be coated with metal
Cannot observe living specimens
Requires vacuum environment