Microscopes

Microscopes are optical instruments that are used in science laboratories to visualize very
minute objects, such as cells and microorganisms, giving a contrasting image that is
magnified.

Microscopes are made up of lenses for magnification, each with its own magnification
powers. Depending on the type of lens, it will magnify the specimen according to its focal
strength.

Their ability to function is because they have been constructed with special components
that enable them to achieve high magnification levels. They can view very small
specimens and distinguish their structural differences, for example, the view of animal and
plant cells viewing microscopic bacterial cells.

There are different types of microscopes like

 light microscope,
 dark-field microscope,
 phase contrast microscope,
 electron microscope,
 fluorescent microscope, etc.

Microscopes are generally made up of structural parts for holding and supporting the
microscope and its components and the optical parts that are used for magnification and
viewing of the specimen images.

Modern microscopes have additional electronics and display devices.

Structural parts of a microscope and their functions

There are three structural parts of the microscope i.e. head, arm, and base.

1. Head – The head is a cylindrical metallic tube that holds the eyepiece lens at one
end and connects to the nose piece at other end.

It is also called a body tube or eyepiece tube. It connects the eyepiece lens to the
objective lens.
 The light coming from objectives will bend inside this tube. In binocular
microscopes, they are adjustable so that the viewer can adjust the eyepiece for
maximum visualization.

2. Arm – This is the part connecting the base to the head and the eyepiece tube to the
base of the microscope.

It supports the head of the microscope and is also used when carrying the
microscope.

Some high-quality microscopes have an articulated arm with more than one joint,
allowing more movement of the microscopic head for better viewing.

3. Base – The base is the lowermost part of the microscope that supports the entire
microscope structure.

It provides stability for the microscope. Illuminators, light switches, and electrical
wiring systems are fitted in the base.

Optical parts of a microscope and their functions

The optical parts of the microscope are used to view, magnify, and produce an image from
a specimen placed on a slide. These parts include:

1. Eyepiece – The eyepiece (ocular Lens) is closest to the viewer’s eye.

They are located at the top of the microscope. This part is used to look at the
specimen.

These lenses come in different magnification powers from 5X to 30X, but the most
common ocular lenses are of 10X or 15X magnification.

They magnify the image for the second time.

2. Eyepiece tube – It’s the eyepiece holder. It carries the eyepiece just above the
objective lens.
 In some microscopes, such as the binoculars, the eyepiece tube is flexible and can be
rotated for maximum visualization for variance in distance. For monocular
microscopes, they are none flexible.

3. Diopter Adjustment – Diopter Adjustment is a control knob present only in the

binocular microscope that is used to change focus on one eyepiece.

It is used to correct any difference in vision and compensate for the differences in
vision between the viewer’s two eyes.

4. Nose piece – A nose piece is a movable circular structure that houses all the
objective lenses. It is also called the revolving turret.

It is connected to the body tube and lies just above the stage. It can be rotated
clockwise or counterclockwise to increase or decrease the magnification.

The change in magnification results due to a change in the objective lens.

5. Objective lenses – The objective lens is the lens that is closest to the specimen.
They are fitted on the nosepiece.

A standard microscope has 3 to 4 objective lenses of different magnifying powers:

4X, 10X, 40X, and 100X.

The objective lenses first receive the light transmitted from the specimen and
magnify the image for the first time.

Objective lenses are color-coded and are of different sizes. Size and color depend on
the power of the lens.

The smallest lens is of the lowest power, and gradually, the longest will be of the
highest power.

The high power lenses i.e. 40X and 100X, are retractable, i.e., their end can be
pushed inward.

In most optical microscopes, objective lenses with 100X or more magnification are
of oil immersion type.
6. The Adjustment knobs – Adjustment Knobs are the control knobs used to focus
the microscope on the specimen.

These knobs are of two types;

a. Fine Adjustment Knob: Fine Adjustment Knob is used for fine adjustment.

It is a smaller knob and is used to move the stage up or down very slowly.

The stage covers a very small distance on each rotation of the fine adjustment knob.
It is used to sharpen the image. It is mostly used while viewing under high power.

b. Coarse Adjustment Knob: Coarse Adjustment Knob is used for focusing the
image under low power magnification.

It is a larger knob and is used to move the stage up or down very rapidly. The stage
is raised or lowered rapidly with the help of a coarse adjustment knob.

7. Stage – This is the section in which the specimen is placed for viewing. They have
stage clips that hold the specimen slides in place.

The most common stage is the mechanical stage, which allows the control of the
slides by moving the slides using the mechanical knobs on the stage instead of
moving them manually.

8. Stage Control Knobs – Stage Control Knobs are the control knobs used to move
the stage mechanically.

There are two knobs; one for moving left and right and the other for moving forward
and backward. This will move the slide in the field of vision.

9. Aperture – This is a hole in the microscope stage through which the transmitted
light from the source reaches the stage.

10. Microscopic illuminator – A microscopic illuminator is a light source. In some

compound microscopes, a mirror, which reflects the light from an external source to
the sample, is used. In other optical microscopes, different electric bulbs of low
voltages are used as a constant light source.
 Commonly used illuminators are tungsten-halogen lamps, 75-150W Xenon lamps,
tin-halide lamps, mercury vapor lamps, etc. The selection of types of bulbs is based
on the requirement of intensity and wavelength for illumination.

11. Condenser – These are lenses that are used to collect and focus light from the
illuminator into the specimen.

They are found under the stage next to the diaphragm of the microscope. They play
a major role in ensuring clear, sharp images are produced with a high magnification
of 400X and above.

The higher the magnification of the condenser, the clearer the image. More
sophisticated microscopes come with an Abbe condenser that has a high
magnification of about 1000X.

12. Diaphragm – It’s also known as the iris. It is found under the stage of the
microscope, and its primary role is to control the amount of light that reaches the
specimen.

It’s an adjustable apparatus, hence controlling the light intensity and the size of the
beam of light that gets to the specimen.

For high-quality microscopes, the diaphragm comes attached with a condenser, and
combined, they are able to control the light focus and light intensity that reaches the
specimen.

13. Condenser focus knob – This is a knob that moves the condenser up or down, thus
controlling the focus of light on the specimen.

14. Brightness Adjustment – The brightness adjustment system controls the voltage
supplied to the light bulb, controlling the intensity (brightness) of the light bulb.

How Does Microscope Work?

In a simple light microscope, a thin specimen containing a slide is placed on the

microscope’s stage.
A beam of light is passed through the condenser to the specimen. The light transmitted
from the specimen enters the objective lens. While passing through the objectives, the
transmitted rays are spread so that they appear to come from the bigger objects.

The light is then focused on the eyepiece lens. This lens further magnifies the pre-
magnified image coming from the objectives.

Finally, a highly magnified image can be observed.

The typical compound light microscope is able to magnify from 40x to 1000x, increasing
our ability to see detail so that objects as small as 0.2 micrometers (μm) or 200 nanometers
(nm) can be seen.

Compound light microscopes may be monocular (for viewing using only one eye) or
binocular (for viewing using both eyes).

Compound light microscopes from various manufacturers may appear different but operate
on similar principles.

A typical school microscope has three magnifications:

Scanning,

Low, and

High dry.

Each objective and eyepiece (ocular lens) will have the magnification written on it. Some
microscopes will also have an oil immersion objective.

The total magnification is the ocular magnification multiplied by the objective

magnification.

Objective Magnification Ocular lens Total magnification

Scanning 4x 10x 40x
Low power 10x 10x 100x
High dry power 40x 10x 400x
Oil immersion 100x 10x 1000x

How to Focus Your Microscope:

The proper way to focus a microscope is to start with the lowest power objective lens first
and while looking from the side, crank the lens down as close to the specimen as possible
without touching it.

Now, look through the eyepiece lens and focus upward only until the image is sharp. If
you can’t get it in focus, repeat the process again.

Once the image is sharp with the low power lens, you should be able to simply click in the
next power lens and do minor adjustments with the fine adjustment knob.
If your microscope has a fine focus adjustment, turning it a bit should be all that’s
necessary. Continue with subsequent objective lenses and fine focus each time.

Note: Both eyes should be open when viewing through the microscope. This prevents eye
fatigue, which occurs when the non-viewing eye is kept closed.

Keeping both eyes open does take some practice, but it is highly recommended. Also, you
should never let your eye touch the ocular lens.

If your eyelashes touch the lens you are to close. Always remove eyeglasses when viewing
through a microscope. If your eyeglass lens touches the microscope it may get scratched.

Rules for Microscope Use:

1. Carry the microscope properly.

2. Always begin focusing with the 4X objective.

3. Use the coarse focus only with the 4X objective in place.

4. Use immersion oil only with the 100X objective (oil immersion lens) in place.

5. Use only ONE drop of oil.

6. Lower the stage and then remove the slide when you are done.

7. ALWAYS clean the microscope when you are done. Use a clean cotton cloth or lens
paper and the alcohol in the labeled jars.

8. Always place the 4X objective over the stage and be sure the stage is at its lowest
position before putting the microscope away.

9. Always turn off the light before putting the microscope away.

10. Always wrap the cord correctly before putting the microscope away.

11. Always return the microscope to the correct cabinet.

12. Always place the oculars toward the BACK of the cabinet.
Cleaning the Microscope

Before using the microscope, it’s essential to clean the lenses and other surfaces to remove
any dust, fingerprints, or debris that may affect image quality. Use a soft, lint-free cloth or
lens paper to gently wipe the lenses. Avoid using harsh chemicals or solvents that may
damage the lens coatings.

Proper Storage

When not in use, store the microscope in a clean and dry environment, preferably in a
dust-free case or cabinet. Protect it from excessive heat, humidity, and direct sunlight, as
these factors can damage the optics and other sensitive components.

Handling with Care

Handle the microscope with care to avoid accidental damage. Always grasp the
microscope by the arm and base, and avoid applying excessive force or pressure on the
lenses or other delicate parts. When moving the microscope, hold it close to your body to
ensure stability.

Microscope Maintenance

Regular maintenance is essential for keeping your microscope in optimal condition.

Follow these guidelines to ensure its longevity and performance.

Regular Cleaning and Maintenance

Periodically clean the microscope using a soft brush or compressed air to remove dust and
debris from the surfaces and crevices. Inspect the microscope for any loose screws or
damaged components and tighten or repair them as necessary. Regularly check the
electrical connections and cables for any signs of wear or damage.

Lens Care

The lenses are critical components of the microscope, and proper care is crucial for
obtaining clear and accurate images. Clean the lenses using a lens cleaning solution and
lens paper or a microfiber cloth. Gently wipe the lenses in a circular motion, starting from
the center and moving outward. Avoid touching the lenses with your fingers to prevent oil
and dirt transfer.
Light Source Care

If your microscope has a built-in light source, such as an LED or halogen lamp, ensure that
it is functioning properly. Periodically check the light intensity and replace the bulb if it’s
dim or flickering. Avoid leaving the light source on for extended periods when not in use,
as it can cause overheating and reduce its lifespan.

Adjusting the Microscope

Proper adjustment of the microscope is essential for obtaining clear and focused images.
Here are the key adjustments you need to consider.

Focusing the Microscope

To bring the specimen into focus, start with the lowest magnification objective lens and
use the coarse focus adjustment knob to get a rough focus. Once the image is roughly in
focus, use the fine focus adjustment knob for precise focusing. Repeat this process when
switching to higher magnification objectives.

Adjusting Magnification

Microscopes often have multiple objective lenses with different magnification levels. To
change the magnification, rotate the nosepiece (also known as the turret) to select the
desired objective lens. Make sure to refocus the image after changing the magnification.

Sample Preparation

Before placing a sample on the microscope, it needs to be properly prepared to ensure

clear visibility and accurate observation. Follow these steps for sample preparation.

Preparing Slides

1. Clean the microscope slides to remove any dirt or residue.

2. Place the specimen on the center of the slide.
3. Add a drop of mounting medium or a suitable liquid (e.g., water, immersion oil) to
the specimen.
4. Carefully place a coverslip over the specimen, starting from one edge and gently
lowering it to avoid trapping air bubbles.
Using Coverslips

Coverslips protect the specimen and ensure a flat surface for observation. Make sure the
coverslip is clean and free from any smudges or debris before placing it on the specimen.
Avoid pressing too hard to prevent damage to the slide or specimen.

Illumination Techniques

Microscopes employ different illumination techniques to enhance the contrast and

visibility of the specimen. Here are some commonly used techniques:

Brightfield Illumination

This is the standard illumination technique where the specimen is illuminated from below.
It is suitable for observing stained or naturally pigmented samples.

Darkfield Illumination

Darkfield illumination involves illuminating the specimen at an oblique angle, resulting in

a dark background. It is useful for observing transparent or unstained specimens with low
contrast.

Phase Contrast Illumination

Phase contrast microscopy is ideal for observing living cells and transparent specimens. It
enhances the contrast by exploiting differences in refractive index within the specimen.

Fluorescence Illumination

Fluorescence microscopy uses fluorescent dyes or labels to visualize specific structures or

molecules within the specimen. It requires a specialized microscope equipped with
appropriate filters and a light source.

Capturing Images and Videos

Advancements in technology have made it possible to capture high-quality images and

videos of the microscopic world. Here are some methods to consider:
Digital Imaging

Modern microscopes often come with built-in digital cameras or the ability to attach
external cameras. These cameras capture the image directly into a computer or other
digital devices, allowing for easy storage and analysis.

Camera Attachments

If your microscope doesn’t have a built-in camera, you can attach an external camera to
the eyepiece using adapters. These cameras range from basic models to advanced ones
equipped with specialized features for microscopy.

Image and Video Processing

After capturing the images or videos, you can enhance them using image processing
software. Adjusting brightness, contrast, and sharpness can improve the quality and clarity
of the visuals.

Best Practices for Microscope Use

To ensure accurate observations and reliable results, it’s important to follow these best
practices when using a microscope:

Using Proper Lighting

Adjust the intensity of the light source to ensure optimal illumination without causing
glare or overheating. The appropriate lighting conditions can enhance visibility and
contrast.

Controlling Environmental Factors

Maintain a controlled environment when using the microscope. Factors such as

temperature, humidity, and vibrations can affect the quality of observations. Keep the
microscope in a stable and suitable environment to minimize these effects.
Avoiding Contamination

Prevent contamination of the microscope and your samples by practicing good laboratory
hygiene. Clean the stage and other surfaces between samples, use clean tools, and avoid
touching the lenses or other optical surfaces with bare hands.

Safety Precautions

When working with a microscope, it’s important to prioritize safety. Consider the
following precautions:

Electrical Safety

Ensure that the microscope is properly grounded and that the electrical cords are in good
condition. Avoid exposing the microscope to liquids or operating it with wet hands to
prevent electrical shocks.

Chemical Safety

When using chemicals for staining or sample preparation, follow proper safety protocols.
Use appropriate protective equipment, such as gloves and goggles, and work in a well-
ventilated area to avoid inhaling hazardous fumes or coming into direct contact with
harmful substances.

Eye and Skin Protection

Wear safety goggles to protect your eyes from potential splashes or spills. Additionally, if
you’re working with chemicals, use gloves to protect your skin from contact with irritants
or corrosive substances.

Troubleshooting Common Issues

Even with proper care and maintenance, microscopes can encounter issues that affect their
performance. Here are some common problems and possible solutions:
Blurry Images

If your images appear blurry, check the focus adjustment and make sure the specimen is
properly mounted. Clean the lenses to remove any smudges or dirt that may be causing
distortion.

Uneven Illumination

Uneven illumination can result from improper alignment of the light source or issues with
the condenser. Adjust the lighting settings and ensure the condenser is correctly
positioned.

Stuck or Damaged Components

If any components of the microscope become stuck or damaged, avoid forcing them.
Consult the microscope’s user manual or seek professional assistance to avoid causing
further damage.

How do I clean the lenses of a microscope?

To clean the lenses of a microscope, use a soft, lint-free cloth or lens paper. Avoid
touching the lenses with your fingers and gently wipe them in a circular motion, starting
from the center and moving outward.

Can I use a microscope without preparing slides?

Yes, you can use a microscope without preparing slides by using a microscope slide with a
coverslip or utilizing specialized slide holders. However, preparing slides allows for better
sample stability and observation.

What is the difference between bright field and dark field illumination?

Bright field illumination involves illuminating the specimen from below, providing a
bright background. Dark field illumination illuminates the specimen at an oblique angle,
resulting in a dark background, which enhances contrast for transparent specimens.

What safety measures should I follow when using a microscope?

Safety notes that should be followed when using a microscope can be summarized as:
 Always carry the microscope with two hands. It is advisable to place one hand under
the base of the box for extra support.
 If the microscope is stored in a box, always ensure the door is locked before picking
up the box.
 If using sunlight as the external light source, never face the adjustable mirror
directly into the sun.
 Never disassemble the microscope as doing so may damage it.
 Unplug from the power supply before replacing the bulb or moving the microscope.
 Never use coarse focus except with the scanning and low power objective lens. It is
very easy to drive an objective through a slide.
 Make sure your workstation is set up ergonomically to use the microscope.
 Avoid prolonged use of the microscope. Take breaks to rest your eyes and make
sure that the light intensity is not excessive.
 Make sure the microscope and any external light sources are regularly electrically
tested and tagged.