Figure 2. [Spectral overlap and FRET. The...]. - Assay Guidance Manual

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Markossian S, Grossman A, Arkin M, et al., editors. Assay Guidance Manual [Internet]. Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences; 2004-.

Assay Guidance Manual [Internet].

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Figure 2.

Spectral overlap and FRET. The donor (Cy3) is excited at a maximal wavelength of 550 nm, and emits with light at 650 nm. The acceptor (Cy5) absorbs light at 570 nm and emits at 670 nm. Energy transfer occurs due to the spectral overlap between the emission of the donor and excitation spectrum of the acceptor (shown in green). Crosstalk occurs when donor emission overlaps with acceptor emission, such that the donor emission is counted as FRET; similarly, the acceptor can be directly excited during donor excitation if their absorbance spectra overlap (as they do here).

From: Inhibition of Protein-Protein Interactions: Cell-Based Assays

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Wade M, Méndez J, Coussens NP, et al. Inhibition of Protein-Protein Interactions: Cell-Based Assays. 2017 Nov 20. In: Markossian S, Grossman A, Arkin M, et al., editors. Assay Guidance Manual [Internet]. Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences; 2004-. Figure 2. [Spectral overlap and FRET. The...].
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