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Table 2 Comparison between BiFC and FRET.

From: The visible touch: in planta visualization of protein-protein interactions by fluorophore-based methods

  BiFC FRET
Required microscopic equipment simple extensive
Data analysis and computation required - +
Concentration dependence high high (channel FRET, FSPIM) or low (DFRAP, FLIM)
Specific problems false positives (possibly due to high expression levels and/or irreversibility) donor bleed-through (channel FRET, FSPIM), photoconversion, protein mobility (DFRAP)
Endogenous expression control (i.e. visualization of tagged partners with subcellular resolution) - +
Monitoring interaction dynamics - (fluorophore reconstitution irreversible) + (interactions reversible)
Subcellular resolution of interaction sites high high (FLIM) or low (channel FRET, FSPIM or DFRAP)
Suitable for tri-molecular interactions - +/-
Suitable for monitoring multiple distinct interaction pairs inside the same cell ("multicolor") + -
Suitable for medium to high throughput + -