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.
Total internal reflection fluorescence microscopeA 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. TIRFM is an imaging modality which uses the excitation of fluorescent cells in a thin optical specimen section that is supported on a glass slide. The technique is based on the principle that when excitation light is totally internally reflected in a transparent solid coverglass at its interface with a liquid medium, an electromagnetic field, also known as an evanescent wave, is generated at the solid-liquid interface with the same frequency as the excitation light.
MicroscopeA 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. There are many types of microscopes, and they may be grouped in different ways.
Point spread functionThe point spread function (PSF) describes the response of a focused optical imaging system to a point source or point object. A more general term for the PSF is the system's impulse response; the PSF is the impulse response or impulse response function (IRF) of a focused optical imaging system. The PSF in many contexts can be thought of as the extended blob in an image that represents a single point object, that is considered as a spatial impulse. In functional terms, it is the spatial domain version (i.e.
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.
Oil immersionIn light microscopy, oil immersion is a technique used to increase the resolving power of a microscope. This is achieved by immersing both the objective lens and the specimen in a transparent oil of high refractive index, thereby increasing the numerical aperture of the objective lens. Without oil, light waves reflect off the slide specimen through the glass cover slip, through the air, and into the microscope lens (see the colored figure to the right).
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.
HolographyHolography is a technique that enables a wavefront to be recorded and later re-constructed. Holography is best known as a method of generating real , but it also has a wide range of other applications. In principle, it is possible to make a hologram for any type of wave. A hologram is made by superimposing a second wavefront (normally called the reference beam) on the wavefront of interest, thereby generating an interference pattern which is recorded on a physical medium.
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.
Objective (optics)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 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. The objective lens of a microscope is the one at the bottom near the sample.
Dark-field microscopyDark-field microscopy (also called dark-ground microscopy) describes microscopy methods, in both light and electron microscopy, which exclude the unscattered beam from the image. As a result, the field around the specimen (i.e., where there is no specimen to scatter the beam) is generally dark. In optical microscopes a darkfield condenser lens must be used, which directs a cone of light away from the objective lens. To maximize the scattered light-gathering power of the objective lens, oil immersion is used and the numerical aperture (NA) of the objective lens must be less than 1.
Near-field scanning optical microscopeNear-field scanning optical microscopy (NSOM) or scanning near-field optical microscopy (SNOM) is a microscopy technique for nanostructure investigation that breaks the far field resolution limit by exploiting the properties of evanescent waves. In SNOM, the excitation laser light is focused through an aperture with a diameter smaller than the excitation wavelength, resulting in an evanescent field (or near-field) on the far side of the aperture.
Optical sectioningOptical 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. Good optical sectioning, often referred to as good depth or z resolution, is popular in modern microscopy as it allows the three-dimensional reconstruction of a sample from images captured at different focal planes.
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.
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.
Inverted microscopeAn inverted microscope is a microscope with its light source and condenser on the top, above the stage pointing down, while the objectives and turret are below the stage pointing up. It was invented in 1850 by J. Lawrence Smith, a faculty member of Tulane University (then named the Medical College of Louisiana). The stage of an inverted microscope is usually fixed, and focus is adjusted by moving the objective lens along a vertical axis to bring it closer to or further from the specimen.
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.
Huygens–Fresnel principleThe Huygens–Fresnel principle (named after Dutch physicist Christiaan Huygens and French physicist Augustin-Jean Fresnel) states that every point on a wavefront is itself the source of spherical wavelets, and the secondary wavelets emanating from different points mutually interfere. The sum of these spherical wavelets forms a new wavefront. As such, the Huygens-Fresnel principle is a method of analysis applied to problems of luminous wave propagation both in the far-field limit and in near-field diffraction as well as reflection.
Wavefront codingIn optics and signal processing, wavefront coding refers to the use of a phase modulating element in conjunction with deconvolution to extend the depth of field of a digital imaging system such as a video camera. Wavefront coding falls under the broad category of computational photography as a technique to enhance the depth of field. The wavefront of a light wave passing through the camera system is modulated using optical elements that introduce a spatially varying optical path length.
Defocus aberrationIn optics, defocus is the aberration in which an image is simply out of focus. This aberration is familiar to anyone who has used a camera, videocamera, microscope, telescope, or binoculars. Optically, defocus refers to a translation of the focus along the optical axis away from the detection surface. In general, defocus reduces the sharpness and contrast of the . What should be sharp, high-contrast edges in a scene become gradual transitions. Fine detail in the scene is blurred or even becomes invisible.
