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Alternatively the intrinsic fluorescence of a sample (i.e., autofluorescence) can be used. [1] In the life sciences fluorescence microscopy is a powerful tool which allows the specific and sensitive staining of a specimen in order to detect the distribution of proteins or other molecules of interest. As a result, there is a diverse range of ...
Through fluorescence polarization microscopy (FPM), the dipole orientation as well as the intensity of fluorescent probes could be measured. Compared with X-ray crystallography or electron microscopy which could elucidate ultra-high resolution of individual proteins or macromolecule assemblies, FPM doesn't require complex sample preparation ...
Permeabilization involves treatment of cells with (usually) a mild surfactant. This treatment dissolves cell membranes, and allows larger dye molecules into the cell's interior. Mounting usually involves attaching the samples to a glass microscope slide for observation and analysis. In some cases, cells may be grown directly on a slide.
In preparation for imaging with a fluorescence microscope, different methods can be used, such as fluorophores or dyes, immunolabeling, and genetically encoded fluorescent proteins. Different fluorescent labels can be used in order to highlight multiple regions of interest in the sample. [ 2 ]
The binding of up to 48 fluorescent labeled oligos to a single molecule of mRNA provides sufficient fluorescence to accurately detect and localize each target mRNA in a wide-field fluorescent microscopy image. Probes not binding to the intended sequence do not achieve sufficient localized fluorescence to be distinguished from background. [18]
Fluorescence-lifetime imaging microscopy or FLIM is an imaging technique based on the differences in the exponential decay rate of the photon emission of a fluorophore from a sample. It can be used as an imaging technique in confocal microscopy , two-photon excitation microscopy , and multiphoton tomography.
Any change of the chemical microenvironment of the fluorescent probe, as well as changes in the hydration shell of biomolecules result in a relative change of the fluorescence detected when a temperature gradient is applied and can be used to determine binding affinities. MST allows measurement of interactions directly in solution without the ...
Electron microscopy is a method that can take a picture of a whole virus and can reveal its shape and structure. It is not typically used as a routine diagnostic test as it requires a highly specialized type of sample preparation, microscope and technical expertise.