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Fluorescence spectroscopy (also known as fluorimetry or spectrofluorometry) is a type of electromagnetic spectroscopy that analyzes fluorescence from a sample. It involves using a beam of light, usually ultraviolet light , that excites the electrons in molecules of certain compounds and causes them to emit light; typically, but not necessarily ...
A simplified Jablonski diagram illustrating the change of energy levels.. The principle behind fluorescence is that the fluorescent moiety contains electrons which can absorb a photon and briefly enter an excited state before either dispersing the energy non-radiatively or emitting it as a photon, but with a lower energy, i.e., at a longer wavelength (wavelength and energy are inversely ...
Fluorescence anisotropy or fluorescence polarization is the phenomenon where the light emitted by a fluorophore has unequal intensities along different axes of polarization. Early pioneers in the field include Aleksander Jablonski , Gregorio Weber , [ 1 ] and Andreas Albrecht. [ 2 ]
Conventional fluorescence microscopy is performed by selectively staining the sample with fluorescent molecules, either linked to antibodies as in immunohistochemistry or using fluorescent proteins genetically fused to the genes of interest. Typically, the more concentrated the fluorophores, the better the contrast of the fluorescence image.
Time-resolved fluorescence (TRF) measurement is very similar to fluorescence intensity (FI) measurement. The only difference is the timing of the excitation/measurement process. When measuring FI, the excitation and emission processes are simultaneous: the light emitted by the sample is measured while excitation is taking place.
Decreased Fluorescence in a Defined region (the red box) Adjacent to a Bleached Region (the circle) Fluorescence Loss in Photobleaching (FLIP) is a fluorescence microscopy technique used to examine movement of molecules inside cells and membranes. A cell membrane is typically labeled with a fluorescent dye to allow for observation.
A filter fluorometer is a type of fluorometer that may be employed in fluorescence spectroscopy. In the fluorometer, a light source emits light of an excitation wavelength that is relevant to the compound to be measured. Filter fluorometers produce specific excitation and emission wavelengths by using optical filters.
A systematic development of the method for the detection of nano-objects started in the early 2000s as a general effort to explore fluorescence-free options for studying single molecules and nano-objects. [2]