Simple Microscope

The simple microscope is generally considered to be the first microscope. It was created in
the 17th century by Antony van Leeuwenhoek, who combined a convex lens with a holder
for specimens. Magnifying between 200 and 300 times, it was essentially a magnifying
glass. While this microscope was simple, it was still powerful enough to provide van
Leeuwenhoek information about biological specimens, including the difference in shapes
between red blood cells. Today, simple microscopes are not used often because the
introduction of a second lens led to the more powerful compound microscope.

Compound Microscope

With two lenses, the compound microscope offers better magnification than a simple
microscope; the second lens magnifies the image of the first. Compound microscopes are
bright field microscopes, meaning that the specimen is lit from underneath, and they can be
binocular or monocular. These devices provide a magnification of 1,000 times, which is
considered to be high, although the resolution is low. This high magnification, however,
allows users to take a close look at objects too small to be seen with the naked eye,
including individual cells. Specimens are usually small and have some degree of
transparency. Because compound microscopes are relatively inexpensive yet useful, they
are used everywhere from research labs to high school biology classrooms

Characteristics Simple Microscope Compound Microscope

Number of lenses One to magnify objects 3-5 to magnify objects

Condenser lens is absent in Condenser lens is present in Compound

Condenser lens
Simple microscope. Microscope.

One side is plain and the other side is

Mirror type Concave reflecting
concave

Source of Light Natural Illuminator

Level of
One level only Higher-level
magnification

Stand is small, hollow cylindrical

Arm is curved and is used to hold the
Structure attached to the base and is used
microscope.
to hold the microscope.

The use of knobs is much, which help in

Usage of course, The usage of course, hooks, and
focusing and as a result, a clear and
hooks, and knobs knobs is not that much.
concise image is seen.

Has a wide range of use such as in

Can only be used in simple ways
studying the structure of different
Used such as enlarging small letters
objects, e.g. details of cells in living
while reading.
organisms.

Magnifying power Up to 300X 2,000X

Adjusting
No Yes
Magnification

BRIGHT FIELD

Brightfield Microscope is also known as the Compound Light Microscope. It is an optical

microscope that uses light rays to produce a dark image against a bright background. It is
the standard microscope that is used in Biology, Cellular Biology, and Microbiological
Laboratory studies.
This microscope is used to view fixed and live specimens, that have been stained with
basic stains which gives a contrast between the image and the image background. It is
specially designed with magnifying glasses known as lenses that modify the specimen to
produce an image seen through the eyepiece.

DARK FIELD

 Microbiology, the branch of science that has so vastly extended and expanded our knowledge of the
living world, owes its existence to Antoni van Leeuwenhoek.
 In 1673, with the aid of a crude microscope consisting of a biconcave lens enclosed in two metal
plates, Leeuwenhoek introduced the world to the existence of microbial forms of life.
 Over the years, microscopes have evolved from the simple, single-lens instrument of Leeuwenhoek,
with a magnification of 300 X, to the present-day electron microscopes capable of magnifications
greater than 250,000X.
 Microscopes are designated as either light microscopes or electron microscopes.
 Light microscopes use visible light or ultraviolet rays to illuminate specimens. They
include brightfield, darkfield, phase-contrast, and fluorescent instruments.
 This is similar to the ordinary light microscope; however, the condenser system is modified so that
the specimen is not illuminated directly.
 The condenser directs the light obliquely so that the light is deflected or scattered from the
specimen, which then appears bright against a dark background.
 Living specimens may be observed more readily with darkfield than with brightfield microscopy.
The dark ground microscopy has the following uses:
 It is useful for the demonstration of very thin bacteria not visible under ordinary illumination since
the reflection of the light makes them appear larger.
 This is a frequently used method for rapid demonstration of Treponema pallidum in clinical
specimens.
 It is also useful for the demonstration of the motility of flagellated bacteria and protozoa.
 Darkfield is used to study marine organisms such as algae, plankton, diatoms, insects,
fibers, hairs, yeast and protozoa as well as some minerals and crystals, thin polymers and some
ceramics.
 Darkfield is used to study mounted cells and tissues.
 It is more useful in examining external details, such as outlines, edges, grain boundaries and surface
defects than internal structure.

PHASE CONTRAST

Phase contrast is a light microscopy technique used to enhance the contrast of images of transparent and colourless
specimens. It enables visualisation of cells and cell components that would be difficult to see using an ordinary light
microscope.
As phase contrast microscopy does not require cells to be killed, fixed or stained, the technique enables living cells,
usually in culture, to be visualised in their natural state. This means biological processes can be seen and recorded at
high contrast and specimen detail can be observed. Fluorescence staining can be used in combination with phase
contrast to further improve the visualisation of samples.
Phase contrast is ideal for thinner samples, therefore an inverted microscope system can be used. This provides the
additional advantage of having more working space. Phase contrast can also be installed on upright microscopes.
 If thicker samples need to be visualised in high-resolution, differential interference contrast (DIC) is a more suitable
technique to use.

Applications of phase contrast

Phase contrast is used to visualise transparent specimens, when high-resolution is not required, including:
 Living cells (usually in culture)
 Microorganisms
 Thin tissue slices
 Fibres
 Subcellular particles, including organelles

Fluorescence Microscope
Microscopes can be used to view different aspects of a sample by varying
how an image is created. Fluorescence microscopes use specific colors of
light to interact with dyes. As the dyes become illuminated, certain structures
can be isolated and viewed with their respective dyes. This type of
microscope is useful to observe specific proteins within a cell. A camera is
usually attached to capture images from the microscope.

Transmission Electron Microscope (TEM)

Like the scanning electron microscope, the transmission electron
microscope (TEM) uses electrons in creating a magnified image, and
samples are scanned in a vacuum so they must be specially prepared.
Unlike the SEM, however, the TEM uses a slide preparation to obtain a 2-D
view of specimens, so it's more suited for viewing objects with some degree
of transparency. A TEM offers a high degree of both magnification and
resolution, making it useful in the physical and biological sciences,
metallurgy, nanotechnology and forensic analysis.

Scanning Electron Microscope (SEM)

The scanning electron microscope, or SEM, uses electrons rather than light for image formation. Samples are
scanned in vacuum or near-vacuum conditions, so they must be specially prepared by first undergoing
dehydration and then being coated with a thin layer of a conducive material, such as gold. After the item is
prepared and placed in the chamber, the SEM produces a 3-D, black-and-white image on a computer screen.
Offering ample control over the amount of magnification, SEMs are used by researchers in the physical, medical
and biological sciences to examine a range of specimens from insects to bones.

TOTAL MAGNIFICATION OF OBJECTIVE

Low Power Objective (10x)
The low power objective lens has more magnification
power than the scanning objective lens, and it is one of
the most helpful lenses when it comes to observing and
analyzing glass slide samples. The total magnification of
a low power objective lens combined with a 10x
eyepiece lens is 100x magnification, giving you a closer
view of the slide than a scanning objective lens without
getting too close for general viewing purposes.

High Power Objective Lens (40x)

The high-powered objective lens (also called “high dry”
lens) is ideal for observing fine details within a specimen
sample. The total magnification of a high-power
objective lens combined with a 10x eyepiece is equal to
400x magnification, giving you a very detailed picture of
the specimen in your slide.

Oil Immersion Objective Lens (100x)

The oil immersion objective lens provides the most
powerful magnification, with a whopping magnification
total of 1000x when combined with a 10x eyepiece. But
the refractive index of air and your glass slide are
slightly different, so a special immersion oil must be
used to help bridge the gap. Without adding a drop
of immersion oil, the oil immersion objective lens will not
function correctly, the specimen will appear blurry, and
you will not achieve an ideal magnification or resolution.
Oil immersion lenses are also available from some
manufacturers in lower magnifications, and provide
higher resolution than their "high dry" counterparts.