Energy transfer fluorescence measurements

Gu, Y., Di, W. L., Kelsell, D. P. and Zicha, D. (2004). Quantitative fluorescence resonance energy transfer (FRET) measurement with acceptor photobleaching and spectral unmixing. J. Microsc. 215, 162-73. 

Van Munster, E. B., Kremers, G. J., Adjobo-Hermans, M. J. and Gadella, T. W., Jr. (2005). Fluorescence resonance energy transfer (FRET) measurement by gradual acceptor photobleaching. J. Microsc. 218, 253-62. 

Adkins, C. E., Pillai, G. V., Kerby, J., et al. (2001) alpha4beta3delta GABA(A) receptors characterized by fluorescence resonance energy transfer-derived measurements of membrane potential. J. Biol. Chem. 276, 38934-38939. 

TIRF can be combined with fluorescence energy transfer to measure distances between fluorophores on a surface in the presence of a large background of bulk-dissolved fluorophores. 

SH3) domain of the Abelson protein tyrosine kinase (c-Abl-SH3) and its known polyproline ligand 3BP2. The insertion of two fluorophores into the ligand sequence gave the possibility to screen the protein communication by fluorescence resonance energy transfer (FRET) measurement. 

This chapter reviews several techniques which combine the use of laser microbeams with antibodies to study molecular and cellular biology. An overview of the basic properties of lasers and their integration with microscopes and computers is provided. Biophysical applications, such as fluorescence recovery after photobleaching to measure molecular mobility and fluorescence resonance energy transfer to measure molecular distances, as well as ablative applications for the selective inactivation of proteins or the selective killing of cells are described. Other techniques, such as optical trapping, that do not rely on the interaction of the laser with the targeting antibody, are also discussed. 

The previous discussion and examples emphasized the use of steady-state fluorescence data. Steady-state data are measured with constant illumination of the sample. The timescale of these measurements is slow relative to the fluorescence decay times. Hence, the effects of the time-dependent processes are averaged to yield the average emission spectra, anisotropies, or extents of energy transfer. Each measured steady-state quantity is the average of the time-dependent values of that quantity averaged over the time-dependent decay of the sample. For instance, the... 

Sabanayi am, CR, Fid, JS, and Meller, A, Using fluorescence resonance energy transfer to measure distances along individual DNA molecules Corrections due to nonideal transfer. Journal of Chemical Physics 122 (2005) art. no.-061103. 

Furthermore, energy transfer dynamics measurements between SRIOI and another dye. Acid Blue 1 (ABl), at the H2O-CCI4 or H2O-DCE interface were measured. The fluorescence dynamics are given by... 

To measure the rate and extent of polymer diffusion across a boundary, one needs a source of contrast. Contrast is achieved by labeling the polymer, for example, employing a small amount of deuterated latex in conjunction with small angle neutron scattering experiments, 10 or with fluorescent labels in conjunction with direct non-radiative energy transfer [DET] measurements.il In our laboratory, we prefer the DET methodology because of our ready access to beam time. Thus the latex we prepare are... 

The measurement of fluorescence intensity from a compound containing cliromophores of two spectral types is an example of a system for which it is reasonable to operate witli tire average rates of energy transfer between spectral pools of molecules. Let us consider tire simple case of two spectral pools of donor and acceptor molecules, as illustrated in figure C3.4.2 [18]. The average rate of energy transfer can be calculated as... 

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