MicroscopyMicroscopy is the technical field of using microscopes to view objects and areas of objects that cannot be seen with the naked eye (objects that are not within the resolution range of the normal eye). There are three well-known branches of microscopy: optical, electron, and scanning probe microscopy, along with the emerging field of X-ray microscopy. Optical microscopy and electron microscopy involve the diffraction, reflection, or refraction of electromagnetic radiation/electron beams interacting with the specimen, and the collection of the scattered radiation or another signal in order to create an image.
ApertureIn optics, an aperture is a hole or an opening through which light travels. More specifically, the aperture and focal length of an optical system determine the cone angle of the bundle of rays that come to a focus in the . An optical system typically has many openings or structures that limit the ray bundles (ray bundles are also known as pencils of light). These structures may be the edge of a lens or mirror, or a ring or other fixture that holds an optical element in place, or may be a special element such as a diaphragm placed in the optical path to limit the light admitted by the system.
Bright-field microscopyBright-field microscopy (BF) is the simplest of all the optical microscopy illumination techniques. Sample illumination is transmitted (i.e., illuminated from below and observed from above) 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.
Diaphragm (optics)In optics, a diaphragm is a thin opaque structure with an opening (aperture) at its center. The role of the diaphragm is to stop the passage of light, except for the light passing through the aperture. Thus it is also called a stop (an aperture stop, if it limits the brightness of light reaching the focal plane, or a field stop or flare stop for other uses of diaphragms in lenses). The diaphragm is placed in the light path of a lens or objective, and the size of the aperture regulates the amount of light that passes through the lens.
Optical microscopeThe 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 object is placed on a stage and may be directly viewed through one or two eyepieces on the microscope.
LensA lens is a transmissive optical device that focuses or disperses a light beam by means of refraction. A simple lens consists of a single piece of transparent material, while a compound lens consists of several simple lenses (elements), usually arranged along a common axis. Lenses are made from materials such as glass or plastic and are ground, polished, or molded to the required shape. A lens can focus light to form an , unlike a prism, which refracts light without focusing.
Confocal microscopyConfocal 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 (a process known as optical sectioning) within an object.
Cardinal point (optics)In Gaussian optics, the cardinal points consist of three pairs of points located on the optical axis of a rotationally symmetric, focal, optical system. These are the focal points, the principal points, and the nodal points. For ideal systems, the basic imaging properties such as image size, location, and orientation are completely determined by the locations of the cardinal points; in fact only four points are necessary: the focal points and either the principal or nodal points.
Shutter (photography)In photography, a shutter is a device that allows light to pass for a determined period, exposing photographic film or a photosensitive to light in order to capture a permanent image of a scene. A shutter can also be used to allow pulses of light to pass outwards, as seen in a movie projector or a signal lamp. A shutter of variable speed is used to control exposure time of the film. The shutter is constructed so that it automatically closes after a certain required time interval.
Super-resolution microscopySuper-resolution microscopy is a series of techniques in optical microscopy that allow such images to have resolutions higher than those imposed by the diffraction limit, which is due to the diffraction of light. Super-resolution imaging techniques rely on the near-field (photon-tunneling microscopy as well as those that use the Pendry Superlens and near field scanning optical microscopy) or on the far-field.
Ray (optics)In optics, a ray is an idealized geometrical model of light or other electromagnetic radiation, obtained by choosing a curve that is perpendicular to the wavefronts of the actual light, and that points in the direction of energy flow. Rays are used to model the propagation of light through an optical system, by dividing the real light field up into discrete rays that can be computationally propagated through the system by the techniques of ray tracing. This allows even very complex optical systems to be analyzed mathematically or simulated by computer.
Optical aberrationIn optics, aberration is a property of optical systems, such as lenses, that causes light to be spread out over some region of space rather than focused to a point. Aberrations cause the image formed by a lens to be blurred or distorted, with the nature of the distortion depending on the type of aberration. Aberration can be defined as a departure of the performance of an optical system from the predictions of paraxial optics.
Vergence (optics)In optics, vergence is the angle formed by rays of light that are not perfectly parallel to one another. Rays that move closer to the optical axis as they propagate are said to be converging, while rays that move away from the axis are diverging. These imaginary rays are always perpendicular to the wavefront of the light, thus the vergence of the light is directly related to the radii of curvature of the wavefronts. A convex lens or concave mirror will cause parallel rays to focus, converging toward a point.
WavefrontIn physics, the wavefront of a time-varying wave field is the set (locus) of all points having the same phase. The term is generally meaningful only for fields that, at each point, vary sinusoidally in time with a single temporal frequency (otherwise the phase is not well defined). Wavefronts usually move with time. For waves propagating in a unidimensional medium, the wavefronts are usually single points; they are curves in a two dimensional medium, and surfaces in a three-dimensional one.
F-numberAn f-number is a measure of the light-gathering ability of any optical system like a camera lens or even the human eye. It is calculated by dividing the system's focal length by the diameter of the entrance pupil. The f-number is also known as the focal ratio, f-ratio, or f-stop, and it is key in determining the depth of field, rate of light scattering, and exposure of a photograph. The f-number is dimensionless that is usually expressed using a lower-case hooked f with the format N, where N is the f-number.
Fermat's principleFermat's principle, also known as the principle of least time, is the link between ray optics and wave optics. Fermat's principle states that the path taken by a ray between two given points is the path that can be traveled in the least time. First proposed by the French mathematician Pierre de Fermat in 1662, as a means of explaining the ordinary law of refraction of light (Fig. 1), Fermat's principle was initially controversial because it seemed to ascribe knowledge and intent to nature.
Phase-contrast microscopyNOTOC Phase-contrast microscopy (PCM) is an optical microscopy technique that converts phase shifts in light passing through a transparent specimen to brightness changes in the image. Phase shifts themselves are invisible, but become visible when shown as brightness variations. When light waves travel through a medium other than a vacuum, interaction with the medium causes the wave amplitude and phase to change in a manner dependent on properties of the medium.
RefractionIn physics, refraction is the redirection of a wave as it passes from one medium to another. The redirection can be caused by the wave's change in speed or by a change in the medium. Refraction of light is the most commonly observed phenomenon, but other waves such as sound waves and water waves also experience refraction. How much a wave is refracted is determined by the change in wave speed and the initial direction of wave propagation relative to the direction of change in speed.
OpticsOptics is the branch of physics that studies the behaviour and properties of light, including its interactions with matter and the construction of instruments that use or detect it. Optics usually describes the behaviour of visible, ultraviolet, and infrared light. Because light is an electromagnetic wave, other forms of electromagnetic radiation such as X-rays, microwaves, and radio waves exhibit similar properties.
Digital microscopeA 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.
