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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 an instrument used to see objects that are too small to be seen by the naked eye. Microscopy is the science of investigating small objects and structures using such an instrument. Microscopic means invisible to the eye unless aided by a microscope.
Optics 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.
In optics, the numerical aperture (NA) of an optical system is a dimensionless number that characterizes the range of angles over which the system can accept or emit light. By incorporating index of refraction in its definition, NA has the property that it is constant for a beam as it goes from one material to another, provided there is no refractive power at the interface. The exact definition of the term varies slightly between different areas of optics. Numerical aperture is commonly used in microscopy to describe the acceptance cone of an objective, and in fiber optics, in which it describes the range of angles within which light that is incident on the fiber will be transmitted along it.
The focal length of an optical system is a measure of how strongly the system converges or diverges light; it is the inverse of the system's optical power. A positive focal length indicates that a system converges light, while a negative focal length indicates that the system diverges light. A system with a shorter focal length bends the rays more sharply, bringing them to a focus in a shorter distance or diverging them more quickly. For the special case of a thin lens in air, a positive focal length is the distance over which initially collimated (parallel) rays are brought to a focus, or alternatively a negative focal length indicates how far in front of the lens a point source must be located to form a collimated beam. For more general optical systems, the focal length has no intuitive meaning; it is simply the inverse of the system's optical power.
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.
Angular resolution describes the ability of any image-forming device such as an optical or radio telescope, a microscope, a camera, or an eye, to distinguish small details of an object, thereby making it a major determinant of image resolution. The closely related term spatial resolution refers to the precision of a measurement with respect to space, which is directly connected to angular resolution in imaging instruments.
A refracting telescope is a type of optical telescope that uses a lens as its objective to form an image. The refracting telescope design was originally used in spy glasses and astronomical telescopes but is also used for long focus camera lenses. Although large refracting telescopes were very popular in the second half of the 19th century, for most research purposes the refracting telescope has been superseded by the reflecting telescope which allows larger apertures. A refractor's magnification is calculated by dividing the focal length of the objective lens by that of the eyepiece.
The resolution of an optical imaging system – a microscope, telescope, or camera – can be limited by factors such as imperfections in the lenses or misalignment. However, there is a principal limit to the resolution of any optical system, due to the physics of diffraction. An optical system with resolution performance at the instrument's theoretical limit is said to be diffraction-limited.
In optical engineering, the objective is the optical element that gathers light from the object being observed and focuses the light rays to produce a real image. Objectives can be a single lens or mirror, or combinations of several optical elements. They are used in microscopes, telescopes, cameras, slide projectors, CD players and many other optical instruments. Objectives are also called object lenses, object glasses, or objective glasses.
An optical system with astigmatism is one where rays that propagate in two perpendicular planes have different foci. If an optical system with astigmatism is used to form an image of a cross, the vertical and horizontal lines will be in sharp focus at two different distances. The term comes from the Greek α- (a-) meaning "without" and στίγμα (stigma), "a mark, spot, puncture".
The point spread function (PSF) describes the response of an imaging system to a point source or point object. A more general term for the PSF is a system's impulse response, the PSF being the impulse response of a focused optical system. The PSF in many contexts can be thought of as the extended blob in an image that represents a single point object. In functional terms it is the spatial domain version of the optical transfer function of the imaging system. It is a useful concept in Fourier optics, astronomical imaging, medical imaging, electron microscopy and other imaging techniques such as 3D microscopy and fluorescence microscopy. The degree of spreading (blurring) of the point object is a measure for the quality of an imaging system. In non-coherent imaging systems such as fluorescent microscopes, telescopes or optical microscopes, the image formation process is linear in the image intensity and described by linear system theory. This means that when two objects A and B are imaged simultaneously, the resulting image is equal to the sum of the independently imaged objects. In other words: the imaging of A is unaffected by the imaging of B and vice versa, owing to the non-interacting property of photons. In space-invariant system, i.e. the PSF is the same everywhere in the imaging space, the image of a complex object is then the convolution of the true object and the PSF. However, when the detected light is coherent, image formation is linear in the complex field. The recorded intensity image then can show cancellations or other non-linear effects.
Confocal microscopy, most frequently confocal laser scanning microscopy (CLSM) or laser confocal scanning microscopy (LCSM), 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.
In optics a ray is an idealized model of light, obtained by choosing a line 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. Ray tracing uses approximate solutions to Maxwell's equations that are valid as long as the light waves propagate through and around objects whose dimensions are much greater than the light's wavelength. Ray theory does not describe phenomena such as diffraction, which require wave theory. Some wave phenomena such as interference can be modeled in limited circumstances by adding phase to the ray model.
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. The only ideal system that has been achieved in practice is the plane mirror, however the cardinal points are widely used to approximate the behavior of real optical systems. Cardinal points provide a way to analytically simplify a system with many components, allowing the imaging characteristics of the system to be approximately determined with simple calculations.
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.
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.
Petzval field curvature, named for Joseph Petzval, describes the optical aberration in which a flat object normal to the optical axis cannot be brought properly into focus on a flat image plane.
A condenser is an optical lens which renders a divergent beam from a point source into a parallel or converging beam to illuminate an object.
The pupil function or aperture function describes how a light wave is affected upon transmission through an optical imaging system such as a camera, microscope, or the human eye. More specifically, it is a complex function of the position in the pupil or aperture that indicates the relative change in amplitude and phase of the light wave. Sometimes this function is referred to as the generalized pupil function, in which case pupil function only indicates whether light is transmitted or not. Imperfections in the optics typically have a direct effect on the pupil function, it is therefore an important tool to study optical imaging systems and their performance.
References
- ↑ Judah Levine, University of Colorado Physics 1230: Light and Color: The Field Lens. Fall 2001
- ↑ SPIE Optopedia: Field Lens
- ↑ Riedl, Max (2001). Optical Design Fundamentals for Infrared Systems. Bellingham, Washington: SPIE Press. p. 34. ISBN 0819440515.
- ↑ Igel, Eugene; Kristiansen, Magne (1997). Rotating Mirror Streak and Framing Cameras. Bellingham, Washington: SPIE Optical Engineering Press. p. 1. ISBN 0819424617.
- ↑ Schmahl, G.; Rudolph, D. (1984). X-Ray Microscopy: Proceedings of the International Symposium, Göttingen, Fed. Rep. of Germany, September 14–16, 1983. Berlin: Springer-Verlag. p. 120. ISBN 9783662135471.
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