Fluorescence microscopeA fluorescence microscope is an optical microscope that uses fluorescence instead of, or in addition to, scattering, reflection, and attenuation or absorption, to study the properties of organic or inorganic substances. "Fluorescence microscope" refers to any microscope that uses fluorescence to generate an image, whether it is a simple set up like an epifluorescence microscope or a more complicated design such as a confocal microscope, which uses optical sectioning to get better resolution of the fluorescence image.
Digital image processingDigital image processing is the use of a digital computer to process s through an algorithm. As a subcategory or field of digital signal processing, digital image processing has many advantages over . It allows a much wider range of algorithms to be applied to the input data and can avoid problems such as the build-up of noise and distortion during processing. Since images are defined over two dimensions (perhaps more) digital image processing may be modeled in the form of multidimensional systems.
Green fluorescent proteinThe green fluorescent protein (GFP) is a protein that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. The label GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria and is sometimes called avGFP. However, GFPs have been found in other organisms including corals, sea anemones, zoanithids, copepods and lancelets. The GFP from A. victoria has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm.
Fluorescent tagIn molecular biology and biotechnology, a fluorescent tag, also known as a fluorescent label or fluorescent probe, is a molecule that is attached chemically to aid in the detection of a biomolecule such as a protein, antibody, or amino acid. Generally, fluorescent tagging, or labeling, uses a reactive derivative of a fluorescent molecule known as a fluorophore. The fluorophore selectively binds to a specific region or functional group on the target molecule and can be attached chemically or biologically.
StainingStaining is a technique used to enhance contrast in samples, generally at the microscopic level. Stains and dyes are frequently used in histology (microscopic study of biological tissues), in cytology (microscopic study of cells), and in the medical fields of histopathology, hematology, and cytopathology that focus on the study and diagnoses of diseases at the microscopic level. Stains may be used to define biological tissues (highlighting, for example, muscle fibers or connective tissue), cell populations (classifying different blood cells), or organelles within individual cells.
FluorophoreA fluorophore (or fluorochrome, similarly to a chromophore) is a fluorescent chemical compound that can re-emit light upon light excitation. Fluorophores typically contain several combined aromatic groups, or planar or cyclic molecules with several π bonds. Fluorophores are sometimes used alone, as a tracer in fluids, as a dye for staining of certain structures, as a substrate of enzymes, or as a probe or indicator (when its fluorescence is affected by environmental aspects such as polarity or ions).
Digital imagingDigital imaging or digital image acquisition is the creation of a digital representation of the visual characteristics of an object, such as a physical scene or the interior structure of an object. The term is often assumed to imply or include the , , , printing and display of such images. A key advantage of a , versus an analog image such as a film photograph, is the ability to digitally propagate copies of the original subject indefinitely without any loss of image quality.
Image editingImage editing encompasses the processes of altering s, whether they are digital photographs, traditional photo-chemical photographs, or illustrations. Traditional analog image editing is known as photo retouching, using tools such as an airbrush to modify photographs or editing illustrations with any traditional art medium. Graphic software programs, which can be broadly grouped into vector graphics editors, raster graphics editors, and 3D modelers, are the primary tools with which a user may manipulate, enhance, and transform images.
FluorescenceFluorescence is the emission of light by a substance that has absorbed light or other electromagnetic radiation. It is a form of luminescence. In most cases, the emitted light has a longer wavelength, and therefore a lower photon energy, than the absorbed radiation. A perceptible example of fluorescence occurs when the absorbed radiation is in the ultraviolet region of the electromagnetic spectrum (invisible to the human eye), while the emitted light is in the visible region; this gives the fluorescent substance a distinct color that can only be seen when the substance has been exposed to UV light.
Flow cytometryFlow cytometry (FC) is a technique used to detect and measure physical and chemical characteristics of a population of cells or particles. In this process, a sample containing cells or particles is suspended in a fluid and injected into the flow cytometer instrument. The sample is focused to ideally flow one cell at a time through a laser beam, where the light scattered is characteristic to the cells and their components. Cells are often labeled with fluorescent markers so light is absorbed and then emitted in a band of wavelengths.
Digital imageA digital image is an composed of picture elements, also known as pixels, each with finite, discrete quantities of numeric representation for its intensity or gray level that is an output from its two-dimensional functions fed as input by its spatial coordinates denoted with x, y on the x-axis and y-axis, respectively. Depending on whether the is fixed, it may be of vector or raster type. Raster image Raster images have a finite set of digital values, called picture elements or pixels.
Fluorescent lampA fluorescent lamp, or fluorescent tube, is a low-pressure mercury-vapor gas-discharge lamp that uses fluorescence to produce visible light. An electric current in the gas excites mercury vapor, which produces short-wave ultraviolet light that then causes a phosphor coating on the inside of the lamp to glow. A fluorescent lamp converts electrical energy into useful light much more efficiently than an incandescent lamp. The typical luminous efficacy of fluorescent lighting systems is 50–100 lumens per watt, several times the efficacy of incandescent bulbs with comparable light output.
Hoechst stainHoechst stains are part of a family of blue fluorescent dyes used to stain DNA. These bis-benzimides were originally developed by Hoechst AG, which numbered all their compounds so that the dye Hoechst 33342 is the 33,342nd compound made by the company. There are three related Hoechst stains: Hoechst 33258, Hoechst 33342, and Hoechst 34580. The dyes Hoechst 33258 and Hoechst 33342 are the ones most commonly used and they have similar excitation–emission spectra.
ImageAn image is a visual representation of something. An image can be a two-dimensional (2D) representation, such as a drawing, painting, or photograph, or a three-dimensional (3D) object, such as a carving or sculpture. An image may be displayed through other media, including projection on a surface, activation of electronic signals, or digital displays. Two-dimensional images can be still or animated. Still images can usually be reproduced through mechanical means, such as photography, printmaking or photocopying.
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.
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.
Speech processingSpeech processing is the study of speech signals and the processing methods of signals. The signals are usually processed in a digital representation, so speech processing can be regarded as a special case of digital signal processing, applied to speech signals. Aspects of speech processing includes the acquisition, manipulation, storage, transfer and output of speech signals. Different speech processing tasks include speech recognition, speech synthesis, speaker diarization, speech enhancement, speaker recognition, etc.
Digital signal processingDigital signal processing (DSP) is the use of digital processing, such as by computers or more specialized digital signal processors, to perform a wide variety of signal processing operations. The digital signals processed in this manner are a sequence of numbers that represent samples of a continuous variable in a domain such as time, space, or frequency. In digital electronics, a digital signal is represented as a pulse train, which is typically generated by the switching of a transistor.
ImagingImaging is the representation or reproduction of an object's form; especially a visual representation (i.e., the formation of an ). Imaging technology is the application of materials and methods to create, preserve, or duplicate images. Imaging science is a multidisciplinary field concerned with the generation, collection, duplication, analysis, modification, and visualization of images, including imaging things that the human eye cannot detect.
Signal processingSignal processing is an electrical engineering subfield that focuses on analyzing, modifying and synthesizing signals, such as sound, , potential fields, seismic signals, altimetry processing, and scientific measurements. Signal processing techniques are used to optimize transmissions, digital storage efficiency, correcting distorted signals, subjective video quality and to also detect or pinpoint components of interest in a measured signal. According to Alan V. Oppenheim and Ronald W.
