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.
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. It is used in optics applied to light waves, in antenna theory applied to radio waves, and in acoustics applied to sound waves. The colloquial use of the term "resolution" sometimes causes confusion; when an optical system is said to have a high resolution or high angular resolution, it means that the perceived distance, or actual angular distance, between resolved neighboring objects is small. The value that quantifies this property, θ, which is given by the Rayleigh criterion, is low for a system with a high 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. The Rayleigh criterion shows that the minimum angular spread that can be resolved by an image-forming system is limited by diffraction to the ratio of the wavelength of the waves to the aperture width. For this reason, high-resolution imaging systems such as astronomical telescopes, long distance telephoto camera lenses and radio telescopes have large apertures.
The 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 shapeless blob in an image that should represent a single point object. We can consider this as a spatial impulse response function. In functional terms, it is the spatial domain version of the optical transfer function (OTF) of an 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.
An X-ray microscope uses electromagnetic radiation in the X-ray band to produce magnified images of objects. Since X-rays penetrate most objects, there is no need to specially prepare them for X-ray microscopy observations.
A superlens, or super lens, is a lens which uses metamaterials to go beyond the diffraction limit. The diffraction limit is a feature of conventional lenses and microscopes that limits the fineness of their resolution depending on the illumination wavelength and the numerical aperture (NA) of the objective lens. Many lens designs have been proposed that go beyond the diffraction limit in some way, but constraints and obstacles face each of them.
Near-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 on the far side of the aperture. When the sample is scanned at a small distance below the aperture, the optical resolution of transmitted or reflected light is limited only by the diameter of the aperture. In particular, lateral resolution of 6 nm and vertical resolution of 2–5 nm have been demonstrated.
Nanophotonics or nano-optics is the study of the behavior of light on the nanometer scale, and of the interaction of nanometer-scale objects with light. It is a branch of optics, optical engineering, electrical engineering, and nanotechnology. It often involves dielectric structures such as nanoantennas, or metallic components, which can transport and focus light via surface plasmon polaritons.
Digital holography is the acquisition and processing of holograms with a digital sensor array, typically a CCD camera or a similar device. Image rendering, or reconstruction of object data is performed numerically from digitized interferograms. Digital holography offers a means of measuring optical phase data and typically delivers three-dimensional surface or optical thickness images. Several recording and processing schemes have been developed to assess optical wave characteristics such as amplitude, phase, and polarization state, which make digital holography a very powerful method for metrology applications .
Vertico spatially modulated illumination (Vertico-SMI) is the fastest light microscope for the 3D analysis of complete cells in the nanometer range. It is based on two technologies developed in 1996, SMI and SPDM. The effective optical resolution of this optical nanoscope has reached the vicinity of 5 nm in 2D and 40 nm in 3D, greatly surpassing the λ/2 resolution limit applying to standard microscopy using transmission or reflection of natural light according to the Abbe resolution limit That limit had been determined by Ernst Abbe in 1873 and governs the achievable resolution limit of microscopes using conventional techniques.
Superoscillation is a phenomenon in which a signal which is globally band-limited can contain local segments that oscillate faster than its fastest Fourier components. The idea is originally attributed to Yakir Aharonov, and has been made more popularly known through the work of Michael Berry, who also notes that a similar result was known to Ingrid Daubechies.
Speckle, speckle pattern, or speckle noise designates the granular structure observed in coherent light, resulting from random interference. Speckle patterns are used in a wide range of metrology techniques, as they generally allow high sensitivity and simple setups. They can also be a limiting factor in imaging systems, such as radar, synthetic aperture radar (SAR), medical ultrasound and optical coherence tomography. Speckle is not external noise; rather, it is an inherent fluctuation in diffuse reflections, because the scatterers are not identical for each cell, and the coherent illumination wave is highly sensitive to small variations in phase changes.
Super-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 or on the far-field. Among techniques that rely on the latter are those that improve the resolution only modestly beyond the diffraction-limit, such as confocal microscopy with closed pinhole or aided by computational methods such as deconvolution or detector-based pixel reassignment, the 4Pi microscope, and structured-illumination microscopy technologies such as SIM and SMI.
The technique of vibrational analysis with scanning probe microscopy allows probing vibrational properties of materials at the submicrometer scale, and even of individual molecules. This is accomplished by integrating scanning probe microscopy (SPM) and vibrational spectroscopy. This combination allows for much higher spatial resolution than can be achieved with conventional Raman/FTIR instrumentation. The technique is also nondestructive, requires non-extensive sample preparation, and provides more contrast such as intensity contrast, polarization contrast and wavelength contrast, as well as providing specific chemical information and topography images simultaneously.
Photo-activated localization microscopy and stochastic optical reconstruction microscopy (STORM) are widefield fluorescence microscopy imaging methods that allow obtaining images with a resolution beyond the diffraction limit. The methods were proposed in 2006 in the wake of a general emergence of optical super-resolution microscopy methods, and were featured as Methods of the Year for 2008 by the Nature Methods journal. The development of PALM as a targeted biophysical imaging method was largely prompted by the discovery of new species and the engineering of mutants of fluorescent proteins displaying a controllable photochromism, such as photo-activatible GFP. However, the concomitant development of STORM, sharing the same fundamental principle, originally made use of paired cyanine dyes. One molecule of the pair, when excited near its absorption maximum, serves to reactivate the other molecule to the fluorescent state.
Light sheet fluorescence microscopy (LSFM) is a fluorescence microscopy technique with an intermediate-to-high optical resolution, but good optical sectioning capabilities and high speed. In contrast to epifluorescence microscopy only a thin slice of the sample is illuminated perpendicularly to the direction of observation. For illumination, a laser light-sheet is used, i.e. a laser beam which is focused only in one direction. A second method uses a circular beam scanned in one direction to create the lightsheet. As only the actually observed section is illuminated, this method reduces the photodamage and stress induced on a living sample. Also the good optical sectioning capability reduces the background signal and thus creates images with higher contrast, comparable to confocal microscopy. Because light sheet fluorescence microscopy scans samples by using a plane of light instead of a point, it can acquire images at speeds 100 to 1,000 times faster than those offered by point-scanning methods.
Super-resolution photoacoustic imaging is a set of techniques used to enhance spatial resolution in photoacoustic imaging. Specifically, these techniques primarily break the optical diffraction limit of the photoacoustic imaging system. It can be achieved in a variety of mechanisms, such as blind structured illumination, multi-speckle illumination, or photo-imprint photoacoustic microscopy in Figure 1.
A virtually imaged phased array (VIPA) is an angular dispersive device that, like a prism or a diffraction grating, splits light into its spectral components. The device works almost independently of polarization. In contrast to prisms or regular diffraction gratings, the VIPA has a much higher angular dispersion but has a smaller free spectral range. This aspect is similar to that of an Echelle grating, since it also uses high diffraction orders. To overcome this disadvantage, the VIPA can be combined with a diffraction grating. The VIPA is a compact spectral disperser with high wavelength resolving power.
Deep learning in photoacoustic imaging combines the hybrid imaging modality of photoacoustic imaging (PA) with the rapidly evolving field of deep learning. Photoacoustic imaging is based on the photoacoustic effect, in which optical absorption causes a rise in temperature, which causes a subsequent rise in pressure via thermo-elastic expansion. This pressure rise propagates through the tissue and is sensed via ultrasonic transducers. Due to the proportionality between the optical absorption, the rise in temperature, and the rise in pressure, the ultrasound pressure wave signal can be used to quantify the original optical energy deposition within the tissue.
Video super-resolution (VSR) is the process of generating high-resolution video frames from the given low-resolution video frames. Unlike single-image super-resolution (SISR), the main goal is not only to restore more fine details while saving coarse ones, but also to preserve motion consistency.
Structured illumination light sheet microscopy (SI-LSM) is an optical imaging technique used for achieving volumetric imaging with high temporal and spatial resolution in all three dimensions. It combines the ability of light sheet microscopy to maintain spatial resolution throughout relatively thick samples with the higher axial and spatial resolution characteristic of structured illumination microscopy. SI-LSM can achieve lateral resolution below 100 nm in biological samples hundreds of micrometers thick.
