Microscopy is the technical field of using microscopes to view objects and areas of objects that cannot be seen with the naked eye. There are three well-known branches of microscopy: optical, electron, and scanning probe microscopy, along with the emerging field of X-ray microscopy.
A microscope is a laboratory instrument used to examine objects that are too small to be seen by the naked eye. Microscopy is the science of investigating small objects and structures using a microscope. Microscopic means being invisible to the eye unless aided by a microscope.
A scanning electron microscope (SEM) is a type of electron microscope that produces images of a sample by scanning the surface with a focused beam of electrons. The electrons interact with atoms in the sample, producing various signals that contain information about the surface topography and composition of the sample. The electron beam is scanned in a raster scan pattern, and the position of the beam is combined with the intensity of the detected signal to produce an image. In the most common SEM mode, secondary electrons emitted by atoms excited by the electron beam are detected using a secondary electron detector. The number of secondary electrons that can be detected, and thus the signal intensity, depends, among other things, on specimen topography. Some SEMs can achieve resolutions better than 1 nanometer.
The optical microscope, also referred to as a light microscope, is a type of microscope that commonly uses visible light and a system of lenses to generate magnified images of small objects. Optical microscopes are the oldest design of microscope and were possibly invented in their present compound form in the 17th century. Basic optical microscopes can be very simple, although many complex designs aim to improve resolution and sample contrast.
The microscopic scale is the scale of objects and events smaller than those that can easily be seen by the naked eye, requiring a lens or microscope to see them clearly. In physics, the microscopic scale is sometimes regarded as the scale between the macroscopic scale and the quantum scale. Microscopic units and measurements are used to classify and describe very small objects. One common microscopic length scale unit is the micrometre, which is one millionth of a metre.
In optical engineering, an objective is an optical element that gathers light from an object being observed and focuses the light rays from it to produce a real image of the object. Objectives can be a single lens or mirror, or combinations of several optical elements. They are used in microscopes, binoculars, telescopes, cameras, slide projectors, CD players and many other optical instruments. Objectives are also called object lenses, object glasses, or objective glasses.
An eyepiece, or ocular lens, is a type of lens that is attached to a variety of optical devices such as telescopes and microscopes. It is named because it is usually the lens that is closest to the eye when someone looks through an optical device to observe an object or sample. The objective lens or mirror collects light from an object or sample and brings it to focus creating an image of the object. The eyepiece is placed near the focal point of the objective to magnify this image to the eyes. The amount of magnification depends on the focal length of the eyepiece.
A total internal reflection fluorescence microscope (TIRFM) is a type of microscope with which a thin region of a specimen, usually less than 200 nanometers can be observed.
Confocal microscopy, most frequently confocal laser scanning microscopy (CLSM) or laser scanning confocal microscopy (LSCM), is an optical imaging technique for increasing optical resolution and contrast of a micrograph by means of using a spatial pinhole to block out-of-focus light in image formation. Capturing multiple two-dimensional images at different depths in a sample enables the reconstruction of three-dimensional structures within an object. This technique is used extensively in the scientific and industrial communities and typical applications are in life sciences, semiconductor inspection and materials science.
Metallography is the study of the physical structure and components of metals, by using microscopy.
Dark-field microscopy describes microscopy methods, in both light and electron microscopy, which exclude the unscattered beam from the image. Consequently, the field around the specimen is generally dark.
Bright-field microscopy (BF) is the simplest of all the optical microscopy illumination techniques. Sample illumination is transmitted white light, and contrast in the sample is caused by attenuation of the transmitted light in dense areas of the sample. Bright-field microscopy is the simplest of a range of techniques used for illumination of samples in light microscopes, and its simplicity makes it a popular technique. The typical appearance of a bright-field microscopy image is a dark sample on a bright background, hence the name.
Köhler illumination is a method of specimen illumination used for transmitted and reflected light optical microscopy. Köhler illumination acts to generate an even illumination of the sample and ensures that an image of the illumination source is not visible in the resulting image. Köhler illumination is the predominant technique for sample illumination in modern scientific light microscopy. It requires additional optical elements which are more expensive and may not be present in more basic light microscopes.
Harold Horace Hopkins FRS was a British physicist. His Wave Theory of Aberrations,, is central to all modern optical design and provides the mathematical analysis which enables the use of computers to create the highest quality lenses. In addition to his theoretical work, his many inventions are in daily use throughout the world. These include zoom lenses, coherent fibre-optics and more recently the rod-lens endoscopes which 'opened the door' to modern key-hole surgery. He was the recipient of many of the world's most prestigious awards and was twice nominated for a Nobel Prize. His citation on receiving the Rumford Medal from the Royal Society in 1984 stated: "In recognition of his many contributions to the theory and design of optical instruments, especially of a wide variety of important new medical instruments which have made a major contribution to clinical diagnosis and surgery."
A digital microscope is a variation of a traditional optical microscope that uses optics and a digital camera to output an image to a monitor, sometimes by means of software running on a computer. A digital microscope often has its own in-built LED light source, and differs from an optical microscope in that there is no provision to observe the sample directly through an eyepiece. Since the image is focused on the digital circuit, the entire system is designed for the monitor image. The optics for the human eye are omitted.
Optical sectioning is the process by which a suitably designed microscope can produce clear images of focal planes deep within a thick sample. This is used to reduce the need for thin sectioning using instruments such as the microtome. Many different techniques for optical sectioning are used and several microscopy techniques are specifically designed to improve the quality of optical sectioning.
A USB microscope is a low-powered digital microscope which connects to a computer's USB port. Microscopes essentially the same as USB models are also available with other interfaces either in addition to or instead of USB, such as via WiFi. They are widely available at low cost for use at home or in commerce. Their cost varies in the range of tens to thousands of dollars. In essence, a USB microscope is a webcam with a high-powered macro lens, and generally uses reflected rather than transmitted light, using built-in LED light sources surrounding the lens. The camera is usually sensitive enough not to need additional illumination beyond normal ambient lighting. The camera attaches directly to the USB port of a computer without the need for an eyepiece, and the images are shown directly on the computer's display.
A condenser is an optical lens that renders a divergent light beam from a point light source into a parallel or converging beam to illuminate an object to be imaged.
A macroscope or photomacroscope in its camera-equipped version is a type of optical microscope developed and named by Swiss microscope manufacturers Wild Heerbrugg and later, after that company's merger with Leica in 1987, by Leica Microsystems of Germany, optimised for high quality macro photography and/or viewing using a single objective lens and light path, rather than stereoscopic viewing of specimens, at magnifications up to around x40. The Wild, subsequently Leica "macroscope" line was in production from approximately 1976–2003; it was succeeded by the Leica Z6 and Z16 offerings, which continued an equivalent functionality, but without the "macroscope" designation. The macroscope remains a useful, if somewhat specialised, instrument for examination of relevant specimens in various laboratories today.
A Lieberkühn reflector (also known as Lieberkühn mirror or simply Lieberkühn) is an illumination device for incident light illumination (epi-illumination) in light microscopes. It encircles the objective, with the mirrored surface facing towards the specimen. This allows illuminating an opaque object from the side of the objective, with the light source positioned behind the specimen as in a transmission microscope.
