Microscopes

A microscope is an instrument used to see objects that are too small to be seen by the naked
eye. This is the instrument used in microscopy, which is the science of investigating small
objects and structures using such an instrument. Microscopes are separated into two main
categories, the light and the electron microscopes, but not all are the same, as each one is
built to produce a specific type of image from a specific type of cell.
Firstly, there are the most commonly used, light microscope. Most light microscopes are
compound meaning that they contain at least two lenses. Because of this they magnify
objects in two stages. Light is shone directly onto and through the underside of the
specimen (or is reflected up by a mirror) into the lens which produces the first magnification.
The image produced by the objective lens is then magnified again by the eyepiece lens
which acts as a simple magnifying glass. This magnified image can then be seen by looking
into the eyepiece lens.
Secondly there are electron microscopes. These are less common but have much shorter
wavelengths than light microscopes which allows them to have a much better resolving
power. One type of electron microscope is the scanning electron microscope (SEM), which
are designed to produce images of the surfaces of microscopic objects. At the top of an SEM
there is a powerful electron gun which fires a beam of electrons down into the machine and
onto the specimen. The main part of the machine (where the object is scanned) is contained
within a sealed vacuum chamber because precise electron beams can't travel effectively
through air. Next a positively charged electrode (anode) attracts the electrons and
accelerates them into an energetic beam which is then brought into a precise focus by an
electromagnetic coil, like a lens in a light microscope. Further down the machine another
coil steers the electron beam from side to side, scanning it slowly and methodically across
the specimen’s surface. The electrons effectively bounce back off the specimen after which
they hit a screen creating producing several signals. These signals are then detected by
appropriate detectors, creating an image of the specimen that is highly magnified.
Another type of electron microscope is the transition electron microscope. This is quite like
the light microscope in that the preparation of a thin slice of specimen (100nm thin) is
required. This sample must then be placed in the vacuum chamber located in the middle of
the machine. To the fire the electrons, a high voltage electricity supply powers the cathode,
a heated filament, which fires an electron beam analogous to the beam of light in a light
microscope. The first lens is an electromagnetic coil which concentrates the electrons into a
more powerful beam, with the second lens focussing the beam onto a certain part of the
specimen. The beam passes through the specimen and ‘picks up’ and image of it. The third
and final lens then magnifies the image. This image then becomes visible when the beam of
electrons hits a fluorescent screen at the base of the machine and can be viewed directly
through binoculars at the side or on a TV monitor attached to the image intensifier- which
makes weak images easier to see.
Electron and light microscopes have many differences and similarities. Firstly, an electron
microscope is very expensive to buy and maintain as it requires special environments to
operate in. In addition to this, the electron microscopes require high technical skills to
operate and are therefore limited to necessary specialised research work. A light microscope
on the other hand is significantly cheaper to buy and maintain. Furthermore, they require no
specialised skills to use which is evident through to way they are used regularly in schools.
The most obvious difference between the two types of microscope is that the light
microscope uses a beam of light to gain an image of the specimen while the electron
microscope uses a beam of electrons. These different types of beam then affect whether the
microscope can be used to view dead or living specimens. Light microscopes can view both
dead and living whereas electron microscopes can only view dead specimens under a high
vacuum. This is because electrons are used which are destructive to life. These specimens
can then be viewed through eyepieces with light microscopes but can only be viewed on a
photographic plate or zinc sulphate fluorescent screen with electron microscopes. In
addition to this when magnifying the image, the electron microscope uses magnetic lenses
and the light microscope uses glass lenses. Because of this, light microscopes have a very
low magnification as they only go up to roughly x1,500 and electron microscopes have a very
high magnification of up to x1,000,000.
Another main difference between the two is the resolution. In electron microscopes, the
resolution is roughly 0.01 nanometres while light microscopes have a resolution of
approximately 3 micrometres. Resolution- the level image detailing- can be used in
situations that require greater details such as studying cell nuclei.
While there are multiple advantages to both light microscopes and electron microscopes,
they both possess some limitations that make them unsuitable for certain tasks. With
electron microscopes, the most obvious limitation is that they are very large and expensive.
In addition to this they require special housing and maintenance which can also be very
expensive. Furthermore, samples that can be viewed are limited to those that are electron
transparent, able to tolerate the vacuum chamber and small enough to fit in the chamber.
After this they must undergo a laborious preparation to be viewed in the vacuum chamber.
This then produces an image which is black and white and can only be analysed by those
with special training.
There are also many disadvantages to the light microscope. Firstly, the light microscope has a
significantly lower magnification and resolution than the electron microscopes. The
magnification has a limit of roughly x1500 due to the light’s long wavelength requiring small
spherical lenses to focus and spread the light. The resolution is also a lot lower in light
microscopes than in electron microscopes as the refracted waves of light are spread, blurring
the image. Moreover, viewing and analysing the specimens is very difficult without the use
of dyes. During the dyeing process, living specimens must be killed and fixed, which
eliminates what is normally an advantage for light microscopes.
In conclusion, this shows that light and electron microscopes have a great deal of
differences, but these can be useful for various types of lab work. Electron microscopes are
clearly more complex and produce better quality images, but they do have many limitations
which may make them impractical to use in certain situations. In contrast light microscopes
are simpler and are more practical in the lab, but don’t produce the best images for a more
detailed analysis.