Think of what limitations are put on us by the
restrictions of our own eyesight: things too far away, or
too small, are beyond our ability to observe without some sort of
devices to help. Just as the telescope was a critical device
for astronomers, the microscope was critical to the
progress of biology.
Although magnifying spectacles (glasses, sort of) have been in
widespread use since the 1300s, the use of lenses to see very tiny
objects was a slowly-developing technology. First of all,
the tinier an object is, the less light reflects off it or passes through
it, and
seeing anything really tiny requires a decent illumination system.
Secondly, magnifying lenses used in early microscopes were made of glass
that was not
particularly even or clear, and tended to split light like a
prism, which affected the resolution limits of the lenses.
Resolution can be thought of as the clarity of focus;
technically, it is the limit at which two tiny objects which are
close together stop being visibly separate. The resolution
of early microscopes was very limited by the glass used in the
lenses.
Some important work was done with early
microscopes, although they were primarily more of a toy than a
scientific instrument. Like telescopes that were being
developed through that same period, many early microscopes used lenses
in sequence (making them multi-lens or compound microscopes)
to magnify an image. In 1660, Italian Marcello
Malpighi was able to use a microscope to see blood capillaries in
the tail of a fish, providing powerful supporting evidence that
blood circulates in the body (prior to this, since no one knew of
how blood runs from arteries through capillaries to veins and back, it was
thought that the blood flow was one-way from production in the
intestines to consumption in the body tissues). In 1665,
Englishman Robert Hooke found that cork was full of tiny chambers,
which he called cells (there were no actual cells as they are now
known in the dead cork, but our label did come from his label).
In the 1670s, Dutchman Antony van
Leeuwenhoek, using a special,
especially pure single lens (a simple microscope) placed
in a holder and held up very close to the eye, was the first
person to write extensively about a world of tiny independent
creatures, which he called animalicules, which seemed to exist all around
but were too small to see by eye. Imagine what an odd idea
that must have been to the people of the day!
It wasn't until the 1800s, through a interesting
development period that included instances of thievery, plagiarism and questionable patent
ethics, that many of the distortions of the lenses were corrected.
With the technological limitations for microscopes mostly solved,
by the end of that century microscopes had begun to hit resolution
limits set by physics: to oversimplify, light beams
themselves have a physical size, and when a gap is at or below about 0.2
micrometers the light itself can no longer fit through that gap - you can't see the gap,
just a smudgy blob where the light has been blocked. Objects below that size just could not be resolved
as long as light was used in the imaging system. Light
microscopes (called that because they use light as an imaging
system) would always remain useful, but most of the objects
discussed in the next section on cell structures were invisible to
them. Although there have been some
recent techniques in improving light microscopes' abilities, they
are still limited.
Through the middle of the 1900s, a new system
was developed that could use a beam of electrons, which have an
adjustable beam much smaller than visible light beams. Electron
beams can be narrowed to below the size of atoms.
The beam is focused with magnets and the final image converted to
light in a way similar to how television screens work.
Recent versions of
electron microscopes have been
used to produce images of
molecules and
atoms. Electron
microscopes are more expensive and
complicated than light
microscopes, and the beam needs to travel through a vacuum to
avoid scattering off air atoms. They are very useful in
research but not going to appear in classroom teaching labs any time soon.
There are two basic set-ups that affect
the type of specimen that can be viewed and what the final image
looks like; both set-ups can be found in both light and
electron microscopes. In a typical laboratory class
microscope, light must pass through a specimen to reach the eyes
of the viewer; this set-up is an example of a transmission
microscope. For a specimen to be useful in a
transmission 'scope, the beam needs to be able to pass
through it and reveal its internal details. Specimens may
need to be
stained, colored or darkened with special
dyes, to produce clear details. Colored dyes are used in
light microscopy (the use of microscopes is called
"microscopy"), while
electron microscope stains have heavy
metals in them (the images are
black-and-white, so the stain
just needs to produce variations in electron absorption; colors may
be added later to make some details clearer). Obviously,
large specimens or structures are not going to let the beams
through them; to see large specimens, the original object
must be
sectioned, converted into very thin slices, then stained for detail. Objects are embedded in a
material that
will hold them together (commonly wax for light
microscopy, plastic for electron microscopy) and then thinly
sliced. The ability to translate the two-dimensional
information of sections into three-dimensional concepts is
important in microscopy.
The other set-up involves reflecting the beam
off the surface of the specimen, which creates a much more
three-dimensional image. This is done by
scanning
microscopes, named for the necessity in the electron
version of scanning the surface with the electron beam. A
dissecting microscope is a type of scanning 'scope.