Uses | Small sample observation |
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Notable experiments
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Discovery of cells |
Inventor | Hans Lippershey Zacharias Janssen |
Related items | Optical microscope Electron microscope |
A microscope (from the Greek: μικρός, mikrós, "small" and σκοπεῖν, skopeîn, "to look" or "see") is an instrument to see objects too small for the naked eye. The science of investigating small objects using such an instrument is called microscopy. Microscopic means invisible to the eye unless aided by a microscope.
There are many types of microscopes, the most common and first to be invented is the optical microscope which uses light to image the sample. Other major types of microscopes are the electron microscope (both the transmission electron microscope and the scanning electron microscope) and the various types of scanning probe microscope.
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The first microscope to be developed was the optical microscope, although the original inventor is not easy to identify. An early microscope was made in 1590 in Middelburg, Netherlands.[1] Two eyeglass makers are variously given credit: Hans Lippershey (who developed an early telescope) and Hans Janssen. Giovanni Faber coined the name microscope for Galileo Galilei's compound microscope in 1625 [2] (Galileo had called it the "occhiolino" or "little eye").
The first detailed account of the interior construction of living tissue based on the use of a microscope did not appear until 1644, in Giambattista Odierna's L'ochio della mosca, or The Fly's Eye.[3]
It was not until the 1660s and 1670s that the microscope was used extensively for research in Italy, Holland and England. Marcelo Malpighi in Italy began the analysis of biological structures beginning with the lungs. Robert Hooke's Micrographia had a huge impact, largely because of its impressive illustrations. A greatest contribution came from Antoni van Leeuwenhoek who discovered red blood cells and spermatozoa and helped popularise microscopy as a technique. On 9 October 1676, Leeuwenhoek reported the discovery of micro-organisms.[3]
In 1893 August Köhler development a key technique for sample illumination, Köhler illumination, which is central to modern light microscopy. This method of sample illumination gives rise to extremely even lighting and overcomes many limitations of older techniques of sample illumination. Further developments in sample illumination came from Fritz Zernike in 1953 and George Nomarski 1955 for their development of phase contrast and differential interference contrast illumination which allow imaging of transparent samples.
In the early 1900s a significant alternative to light microscopy was developed, using electrons rather than light to generate the image. Ernst Ruska started development of the first electron microscope in 1931 which was the transmission electron microscope (TEM). The transmission electron microscope works on the same principle as an optical microscope but uses electrons in the place of light and electromagnets in the place of glass lenses. Use of electrons instead of light allows a much higher resolution.
Development of the transmission electron microscope was quickly followed in 1935 by the development of the scanning electron microscope by Max Knoll[4].
Electron microscopes quickly became popular, following the Second World War Ernst Ruska, working at Siemens developed the first commercial transmission electron microscope and major scientific conferences on electron microscopy started being held in the 1950s. In 1965 the first commercial scanning electron microscope was developed by Professor Sir Charles Oatley and his postgraduate student Gary Stewart and marketedCambridge Instrument Company as the "Stereoscan".
"Microscopes" can be separated into optical theory microscopes (Light microscope), electron microscopes (e.g., TEM), and scanning probe microscopes (SPM). Optical microscopes function through the optical theory of lenses in order to magnify the image generated by the passage of a wave through the sample, or reflected by the sample. The waves used are electromagnetic (in optical microscopes) or electron beams (in electron microscopes). Types are the compound light, stereo, and the electronic microscope.
The most common type of microscope—and the first invented—is the optical microscope. This is an optical instrument containing one or more lenses producing an enlarged image of an sample placed in the focal plane. Optical microscopes have refractive glass and occasionally of plastic or quartz, to focus light into the eye or another light detector. Mirror-based optical microscopes operate in the same manner. Typical magnification of a light microscope, assuming visible range light, is up to 1500x with a theoretical resolution limit of around 0.2 micrometres or 200 nanometers. Specialized techniques (e.g., scanning confocal microscopy, Vertico SMI) may exceed this magnification but the resolution is diffraction limited. The use of shorter wavelengths of light, such as the ultraviolet, is one way to improve the spatial resolution of the optical microscope, as are devices such as the near-field scanning optical microscope.
Sarfus, a recent optical technique increases the sensitivity of standard optical microscope to a point it becomes possible to directly visualize nanometric films (down to 0.3 nanometer) and isolated nano-objects (down to 2 nm-diameter). The technique is based on the use of non-reflecting substrates for cross-polarized reflected light microscopy.
Ultraviolet light enables the resolution of microscopic features, as well as to image samples that are transparent to the eye. Near infrared light images circuitry embedded in bonded silicon devices, as silicon is transparent in this region. Many wavelengths of light, ranging from the ultraviolet to the visible are used to excite fluorescence emission from objects for viewing by eye or with sensitive cameras.
Phase contrast microscopy is an optical microscopy illumination technique in which small phase shifts in the light passing through a transparent specimen are converted into amplitude or contrast changes in the image. A phase contrast microscope does not require staining to view the slide. This microscope made it possible to study the cell cycle.
The traditional optical microscope has recently been modified into a digital microscope, where, instead of directly viewing the object, a charge-coupled device (CCD) is used to record the image, which is then displayed on a computer monitor.
Three major variants of electron microscopes exist:
The SEM and STM can also be considered examples of scanning probe microscopy.
Of these techniques AFM and STM are the most commonly used followed by MFM and SNOM/NSOM.
Scanning acoustic microscopes use sound waves to measure variations in acoustic impedance. Similar to Sonar in principle, they are used for such jobs as detecting defects in the subsurfaces of materials including those found in integrated circuits.
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