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An emission map is measured by recording the emission spectra resulting from a range of excitation wavelengths and combining them all together. This is a three dimensional surface data set: emission intensity as a function of excitation and emission wavelengths, and is typically depicted as a contour map.
Emission spectroscopy is a spectroscopic technique which examines the wavelengths of photons emitted by atoms or molecules during their transition from an excited state to a lower energy state. Each element emits a characteristic set of discrete wavelengths according to its electronic structure , and by observing these wavelengths the elemental ...
Wavelengths of maximum absorption (≈ excitation) and emission (for example, Absorption/Emission = 485 nm/517 nm) are the typical terms used to refer to a given fluorophore, but the whole spectrum may be important to consider. The excitation wavelength spectrum may be a very narrow or broader band, or it may be all beyond a cutoff level.
However, according to Kasha's rule, photon emission (termed fluorescence in the case of an S state) is expected in appreciable yield only from the lowest excited state, S 1. Since only one state is expected to yield emission, an equivalent statement of the rule is that the emission wavelength is independent of the excitation wavelength. [3]
Different colors allow for the tracking of different cell types, transcriptional activity, and fusion in proteins. mCherry, out of all of the true monomers developed, has the longest wavelengths, highest photostability, fastest maturation, excellent pH resistance, and is closest to mRFP1 in its excitation and emission maxima. [1]
The acceptor fluorophore in turn emits light at its characteristic wavelength. The two most commonly used lanthanides in life science assays are shown below along with their corresponding acceptor dye as well as their excitation and emission wavelengths and resultant Stokes shift (separation of excitation and emission wavelengths).
Its excitation peak is 513 nm and its emission peak is 527 nm. [2] Like the parent GFP, YFP is a useful tool in cell and molecular biology because the excitation and emission peaks of YFP are distinguishable from GFP which allows for the study of multiple processes/proteins within the same experiment.
When bound to Ca 2+, GCaMP fluoresces green with a peak excitation wavelength of 480 nm and a peak emission wavelength of 510 nm. [3] It is used in biological research to measure intracellular Ca 2+ levels both in vitro and in vivo using virally transfected or transgenic cell and animal lines.