Microscope, instrument that produces enlarged images of small objects, allowing the observer an exceedingly close view of minute structures at a scale convenient for examination and analysis. Although optical microscopes are the subject of this article, an image may also be enlarged by many other wave forms, including acoustic, X-ray, or electron beam, and be received by direct or digital imaging or by a combination of these methods. The microscope may provide a dynamic image (as with conventional optical instruments) or one that is static (as with conventional scanning electron microscopes).
The magnifying power of a microscope is an expression of the number of times the object being examined appears to be enlarged and is a dimensionless ratio. It is usually expressed in the form 10× (for an image magnified 10-fold), sometimes wrongly spoken as “ten eks”—as though the × were an algebraic symbol—rather than the correct form, “ten times.” The resolution of a microscope is a measure of the smallest detail of the object that can be observed. Resolution is expressed in linear units, usually micrometres (μm).
The most familiar type of microscope is the optical, or light, microscope, in which glass lenses are used to form the image. Optical microscopes can be simple, consisting of a single lens, or compound, consisting of several optical components in line. The hand magnifying glass can magnify about 3 to 20×. Single-lensed simple microscopes can magnify up to 300×—and are capable of revealing bacteria—while compound microscopes can magnify up to 2,000×. A simple microscope can resolve below 1 micrometre (μm; one millionth of a metre); a compound microscope can resolve down to about 0.2 μm.
Images of interest can be captured by photography through a microscope, a technique known as photomicrography. From the 19th century this was done with film, but digital imaging is now extensively used instead. Some digital microscopes have dispensed with an eyepiece and provide images directly on the computer screen. This has given rise to a new series of low-cost digital microscopes with a wide range of imaging possibilities, including time-lapse micrography, which has brought previously complex and costly tasks within reach of the young or amateur microscopist.
Other types of microscopes use the wave nature of various physical processes. The most important is the electron microscope, which uses a beam of electrons in its image formation. The transmission electron microscope (TEM) has magnifying powers of more than 1,000,000×. TEMs form images of thin specimens, typically sections, in a near vacuum. A scanning electron microscope (SEM), which creates a reflected image of relief in a contoured specimen, usually has a lower resolution than a TEM but can show solid surfaces in a way that the conventional electron microscope cannot. There are also microscopes that use lasers, sound, or X-rays. The scanning tunneling microscope (STM), which can create images of atoms, and the environmental scanning electron microscope (ESEM), which generates images using electrons of specimens in a gaseous environment, use other physical effects that further extend the types of objects that can be examined.
History Of Optical Microscopes
Three Dutch spectacle makers—Hans Jansen, his son Zacharias Jansen, and Hans Lippershey—have received credit for inventing the compound microscope about 1590. The first portrayal of a microscope was drawn about 1631 in the Netherlands. It was clearly of a compound microscope, with an eyepiece and an objective lens. This kind of instrument, which came to be made of wood and cardboard, often adorned with polished fish skin, became increasingly popular in the mid-17th century and was used by the English natural philosopher Robert Hooke to provide regular demonstrations for the new Royal Society. These demonstrations commenced in 1663, and two years later Hooke published a folio volume titled Micrographia, which introduced a wide range of microscopic views of familiar objects (fleas, lice, and nettles among them). In this book he coined the term cell.
Dutch civil servant Antonie van Leeuwenhoek began his pioneering observations of freshwater microorganisms in the 1670s. He made his postage-stamp-sized microscopes by hand, and the best of them could resolve details around 0.7 μm. His fine specimens discovered in excellent condition at the Royal Society more than three centuries later prove what a great technician he was. Using his simple microscope, Leeuwenhoek effectively launched microbiology in 1674, and single-lensed microscopes remained popular until the 1850s. In 1827 they were used by Scottish botanist Robert Brown to demonstrate the ubiquity of the cell nucleus, a term he coined in 1831.
Simple microscopes using single lenses can generate fine images; however, they can also produce spurious colours due to chromatic aberration, in which different wavelengths of light do not come to the same focus. The aberrations were worse in the compound microscopes of the time, because the lenses magnified the aberrations at least as much as they magnified the images. Although the compound microscopes were beautiful objects that conferred status on their owners, they produced inferior images. In 1733 the amateur English optician Chester Moor Hall found by trial and error that a combination of a convex crown-glass lens and a concave flint-glass lens could help to correct chromatic aberration in a telescope, and in 1774 Benjamin Martin of London produced a pioneering set of colour-corrected lenses for a microscope.
The appearance of new varieties of optical glasses encouraged continued development of the microscope in the 19th century, and considerable improvements were made in understanding the geometric optics of image formation. The concept of an achromatic (non-colour-distorting) microscope objective was finally introduced in 1791 by Dutch optician Francois Beeldsnijder, and the English scientist Joseph Jackson Lister in 1830 published a work describing a theoretical approach to the complete design of microscope objectives. The physics of lens construction was examined by German physicist Ernst Abbe. In 1868 he invented an apochromatic system of lenses, which had even better colour correction than achromatic lenses, and in 1873 he published a comprehensive analysis of lens theory. Light microscopes that were produced in the closing quarter of the 19th century reached the effective limits of optical microscopy. Subsequent instruments, such as phase-contrast microscopes, interference microscopes, and confocal microscopes, solved specific problems that had arisen during the study of specimens such as living cells.
The Simple Microscope
The simple microscope consists of a single lens traditionally called a loupe. The most familiar present-day example is a reading or magnifying glass. Present-day higher-magnification lenses are often made with two glass elements that produce a colour-corrected image.
It is instinctive, when one wishes to examine the details of an object, to bring it as near as possible to the eye. The closer the object is to the eye, the larger the angle that it subtends at the eye, and thus the larger the object appears. If an object is brought too close, however, the eye can no longer form a clear image. The use of the magnifying lens between the observer and the object enables the formation of a “virtual image” that can be viewed in comfort. To obtain the best possible image, the magnifier should be placed directly in front of the eye. The object of interest is then brought toward the eye until a clear image of the object is seen.
The magnifying power, or extent to which the object being viewed appears enlarged, and the field of view, or size of the object that can be viewed, are related by the geometry of the optical system. A working value for the magnifying power of a lens can be found by dividing the least distance of distinct vision by the lens’ focal length, which is the distance from the lens to the plane at which the incoming light is focused. Thus, for example, a lens with a least distance of distinct vision of 25 cm and a focal length of 5 cm (2 inches) will have a magnifying power of about 5×.
The Compound Microscope
The limitations on resolution (and therefore magnifying power) imposed by the constraints of a simple microscope can be overcome by the use of a compound microscope, in which the image is relayed by two lens arrays. One of them, the objective, has a short focal length and is placed close to the object being examined. It is used to form a real image in the front focal plane of the second lens, the eyepiece or ocular. The eyepiece forms an enlarged virtual image that can be viewed by the observer. The magnifying power of the compound microscope is the product of the magnification of the objective lens and that of the eyepiece.