SIMPLE MICROSCOPE

Science behind the history of microscope. In the 17th century when the first microscopes were
invented. There were several epidemic diseases that were poorly understood such as small pox.
Scientists recognized the need for the tool that could help them to better understand the causes and
mechanisms of these diseases and microscope provide them the study of such microorganisms that
could not able to be seen with the naked eye.
A simple microscope is a fundamental optical device comprised of a single lens that magnifies
objects when they are placed close to the lens. This lens, typically convex in shape, bends light
rays passing through it, allowing for the enlargement of the object's image. Simple microscopes
are commonly used for various applications such as reading small text, examining specimens, or
inspecting details in crafts and artwork. They are portable, easy to use, and provide moderate levels
of magnification suitable for everyday tasks requiring enhanced visual clarity.
A simple microscope, also known as an light microscope, consists of several key components that
work together to produce a magnified image of a specimen. Here are the main components.
Mechanical Components:
1. Stage: Holds the specimen in place and allows for precise movement (x-y direction).
2. Stage Clips: Secure the specimen to the stage.
3. Coarse Adjustment Knob: Moves the stage up or down for large-scale focusing.
4. Fine Adjustment Knob: Makes precise adjustments for fine focusing.
5. Eyepiece Lens (Ocular Lens): Magnifies the image formed by the objective lens (typically 10x).
6. Objective Lenses: Collect light from the specimen and form an enlarged image (available in
various magnifications: 4x, 10x, 40x, 100x).
7. Light Source: Built-in (e.g., LED, halogen) or external (e.g., mirror, lamp).
8. Diaphragm: Regulates the amount of light reaching the specimen.
9. Condenser Aperture: Controls the angle of light entering the condenser lens
10. Base: Provides stability and support for the microscope.
11. Arm: Connects the optical components to the base.
12. Eyepiece Holder: Secures the eyepiece lens in place.
Applications of Microscope:
Simple microscope is used to see the objects that is not visible with naked eyes
1- Biology: Studying small plants, animals, microorganism, such as insects, cells, tissues.
2- Giolology: Examining rocks, minerals, and fossils.
3- Lab work: Simple microscope are often used in laboratories for basic experiments.
 4- Jewelry inspection: Jewelry makers can see a magnified image of the small components of
the jewelry by using a simple microscope. Examining gemstones, pearls, and other precious
stones.
5- Forensic science: Examining evidence, such as hair, fibers, and small objects. It is used
during crime scene investigations to separate, distinguish, and compare forensic materials.
6- Dermatology: It is used by dermatologists to research skin diseases. Examining skin
conditions, such as wounds or parasites.
7- Entomology: Studying insects that bite and stings.
8- Food inspection: Checking food samples for contaminants or spoilage.
9- Pharmaceuticals: Inspecting small tablets, capsules, or powders.
Limitations of Simple Microscope
1. Limited magnification: Simple microscopes usually offer low magnification (up to around
10-20x), which is insufficient for observing very small or detailed cellular structures.
2. Low resolution: They have limited resolving power, so they can’t provide clear images of
tiny details compared to compound or electron microscopes.
3. Limited applications: They are mainly suitable for basic observations, so they are not useful
for advanced scientific research for observing microorganism at the cellular level.
4. Manual operation: They usually lack advanced focusing mechanisms, requiring manual
adjustment that can make detailed examination harder.
5. Limited field of view: The field of view is narrow, making it difficult to observe larger
specimens or get a complete picture of certain.
6. Single lens distortion: Simple microscopes uses a single lens, which can lead to optical
distortions like chromatic aberration (color fringing) causing the image to appear blurred
or distorted at the edges.

COMPOUND MICROSCOPE
A Compound Microscope is a type of microscope that uses two or more lenses to magnify small
objects. It typically consists of an objective lenses (near the specimen) and ocular lens (where the
viewer views the image).

• The specimen is mounted on glass slide between condenser and object lenses.
• The condenser lens focuses a beam of visible light on the specimen
• The objective lens creates a magnified image of specimen which is further magnified by
the ocular lenses also known as eyepiece.
• Magnification can be controlled by the rotation of nosepiece.
• For very high magnification (100X), Oil immersion objective lens is used.
 • Light from the light source is diffracted by specimen, collected by Objective lens and
focused on the image plane
• The rays constructively and destructively interfere to form a contrast image.
• The total magnification of Compound microscope depends upon the individual
magnifications of Eyepiece and Objective lens.
• If we have an eyepiece of 10x magnification and an objective lens of 4x magnification
• The total magnification will be: (10x) x (4x) = 40x

Types of Compound Microscope

1. Bright Field Microscope
Brightfield Microscope is an optical microscope that uses light rays to produce a dark image
against a bright background. Brightfield Microscope is also known as the Compound Light
Microscope.
2. Darkfield microscope
A Darkfield microscope is arranged so that the light source is blocked off, causing light to scatter
as it hits the specimen. The design of the dark field microscope is such that it removes the dispersed
light, or zeroth order, so that only the scattered beams hit the sample.
3. Phase-contrast microscopy
Phase-contrast microscopy is an optical microscopy technique that converts phase shifts in the
light passing through a transparent specimen to brightness changes in the image.
4. Polarizing microscope
Polarizing microscope is a type of microscope that uses polarized light to study the optical
properties of materials. It is mainly used in geological studies to study geological specimens. For
this reason, it is also known as a petrographic microscope.

Limitations of Compound Microscope

 As far as the limitations of compound microscope is concerned, the brightness of the image
as seen by our eye depends on how much of our pupil area is filled with light. Now when
you place your eye on the eyepiece of the compound microscope only a small portion of
your pupil will be filled with light.
 Therefore, the level of illumination that you experience while observing the specimen
under a compound microscope is never the same or more than the natural brightness of the
specimen itself.
 It has only one optical path, resulting in a smaller field of view and limited depth of field.
  It cannot observe the deep level structures of specimen. It requires high standards for
specimen preparation.

FLUORESENCE MICROSCOPE
Fluorescence microscopy is a technique that uses fluorescence (the emission of light by a substance
that has absorbed light) to study the properties of organic or inorganic substances.
Basic Principle: A fluorophore absorbs light at one wavelength (excitation) and emits light at a
longer wavelength (emission), which is detected and visualized.
Types of Fluorophores
1. Natural Fluorophores:
• GFP (Green Fluorescent Protein): Derived from jellyfish, used to label live cells.
• Phycoerythrin: Found in algae, useful in flow cytometry.
2. Synthetic Fluorophores:
• DAPI: Binds strongly to DNA and emits blue fluorescence.
• Rhodamine: Red-emitting dye used for imaging organelles.
3. Quantum Dots:
• Semiconductor nanoparticles with high brightness and stability.
Applications of Fluorophores
1. Cell Labeling:
Highlight specific structures (e.g., nuclei, mitochondria).
2. Protein Interaction Studies:
Track protein-protein interactions in live cells.
3. Diagnostic Imaging:
Detect cancer cells, pathogens, or disease markers.
4. Environmental Applications:
Trace pollutants or monitor microbial activity.
Challenges in Fluorescence Microscopy
• High Cost: Advanced equipment and fluorophores are expensive.
• Photobleaching: Fluorophores degrade with prolonged exposure to light.
• Autofluorescence: Background signals can obscure specific fluorescence.
• Complexity: Requires specialized knowledge for setup and interpretation.
• Resolution Limitations: Standard techniques cannot surpass the diffraction limit.