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.
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.
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.
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.
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.
Geometrical opticsGeometrical optics, or ray optics, is a model of optics that describes light propagation in terms of rays. The ray in geometrical optics is an abstraction useful for approximating the paths along which light propagates under certain circumstances. The simplifying assumptions of geometrical optics include that light rays: propagate in straight-line paths as they travel in a homogeneous medium bend, and in particular circumstances may split in two, at the interface between two dissimilar media follow curved paths in a medium in which the refractive index changes may be absorbed or reflected.
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.
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.
Physical opticsIn physics, physical optics, or wave optics, is the branch of optics that studies interference, diffraction, polarization, and other phenomena for which the ray approximation of geometric optics is not valid. This usage tends not to include effects such as quantum noise in optical communication, which is studied in the sub-branch of coherence theory. Physical optics is also the name of an approximation commonly used in optics, electrical engineering and applied physics.
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.
Fourier opticsFourier optics is the study of classical optics using Fourier transforms (FTs), in which the waveform being considered is regarded as made up of a combination, or superposition, of plane waves. It has some parallels to the Huygens–Fresnel principle, in which the wavefront is regarded as being made up of a combination of spherical wavefronts (also called phasefronts) whose sum is the wavefront being studied. A key difference is that Fourier optics considers the plane waves to be natural modes of the propagation medium, as opposed to Huygens–Fresnel, where the spherical waves originate in the physical medium.
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.
Two-photon excitation microscopyTwo-photon excitation microscopy (TPEF or 2PEF) is a fluorescence imaging technique that is particularly well-suited to image scattering living tissue of up to about one millimeter in thickness. Unlike traditional fluorescence microscopy, where the excitation wavelength is shorter than the emission wavelength, two-photon excitation requires simultaneous excitation by two photons with longer wavelength than the emitted light. The laser is focused onto a specific location in the tissue and scanned across the sample to sequentially produce the 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.
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.
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.
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.
Optical discAn optical disc is a flat, usually disc-shaped object that stores information in the form of physical variations on its surface that can be read with the aid of a beam of light. Optical discs can be reflective, where the light source and detector are on the same side of the disc, or transmissive, where light shines through the disc to the be detected on the other side. Optical discs can store analog information (e.g. Laserdisc), digital information (e.g. DVD), or store both on the same disc (e.g. CD Video).
Optical fiberAn optical fiber, or optical fibre in Commonwealth English, is a flexible, transparent fiber made by drawing glass (silica) or plastic to a diameter slightly thicker than that of a human hair. Optical fibers are used most often as a means to transmit light between the two ends of the fiber and find wide usage in fiber-optic communications, where they permit transmission over longer distances and at higher bandwidths (data transfer rates) than electrical cables.
Optical computingOptical computing or photonic computing uses light waves produced by lasers or incoherent sources for data processing, data storage or data communication for computing. For decades, photons have shown promise to enable a higher bandwidth than the electrons used in conventional computers (see optical fibers). Most research projects focus on replacing current computer components with optical equivalents, resulting in an optical digital computer system processing binary data.