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This unspecific background fluorescence can be differentiated from the GFP signal by its spectra and also by its characteristic fluorescence decay pattern.
Over-staining and bright green background fluorescence can be visible and sometimes too high to allow quantification [ 20, 25, 64].
Matthew P (2004) and Liu.W.etc (2002) pointed that background fluorescence can be obtained by the developed four-parametric sigmoid function fit-point method [ 18].
The number of cycles required to produce a signal above background fluorescence can be converted to the starting concentration of the unknowns from the standard curve.
Although some background fluorescence can be observed, we were not able to find a reconstituted YFP signal in these experiments, indicating that the C-terminal hAT blocks D,E and F are indeed responsible for homodimerization.
Replacing the filter-based detectors with a hyperspectral imaging system has been shown to identify and eliminate sources of error, such as spatially localized contaminants [ 1]. Background fluorescence can be removed by post-processing, or reduced via structured illumination of the sample [ 2].
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The advantages of using the FLIM phasor and N&B analyses are that BaP fluorescence can be distinguished from the background fluorescence and aggregation of BaP molecules can be visualized inside the cells.
The fluorescence can be turned on by exposure to visible light, and turned off by exposure to ultraviolet.
Whereas GFP fluorescence shows a prototypical monoexponential decay function, background fluorescence can only be described by a multiexponential decay function.
The chlorophyll fluorescence lifetime can be estimated from deconvolution of the fluorescence and background signals.
This brings the two fluorescent proteins near each other, causing fluorescence that can be seen with a microscope.
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