Fluorescence Resonance Energy Transfer (FRET) Service

CD BioSciences provides clients with fluorescence resonance energy transfer (FRET) service, an advanced technique for investigating various biological phenomena that produce changes in molecular proximity. Our service enables the co-localization of proteins and other molecules to be imaged with spatial resolution beyond the limitations of conventional light microscopy.

FRET describes the mechanism of energy transfer between two chromophores. A donor chromophore in an electronically excited state can transfer energy to an acceptor chromophore through non-radiative dipole-dipole coupling. The energy transfer efficiency is proportional to the distance between the donor and acceptor, making FRET extremely sensitive to small changes in distance. Therefore, FRST can serve as an excellent reporter of molecular proximity and protein interactions.

Figure 1. The mechanism of fluorescence resonance energy transfer (FRET). (Wu, L., et al., 2020)

Service Content

Experimental content

• Cultivate specific cells for transfection according to experimental requirements.
• Construct plasmid vectors donor-protein A and acceptor-protein B.
• Co-transfect cells with both plasmids.
• FRET assay (whether it occurs or not).
• FRET image acquisition.

a) Determine excitation wavelength for FRET pairing.

a) Acquire donor and acceptor images.

a) Collect FRET signal.

• FRET data processing.

a) Removal of spectral crosstalk from FRET signals.

b) Correct for changes in spectral sensitivity of the FRET signal in the acceptor pathway.

c) Correction for autofluorescence and optical noise.

d) Pixel matching correction for dual-calibration cells using correction coefficients.

Assay FRET efficiency service

FRET is effective in quantifying molecular dynamics in plant protein-protein interactions. The energy transfer efficiency between donors and acceptors is proportional to their distance. Therefore, assay FRET efficiency can identify interactions between labeled complexes.

• Photobleaching FRET

We infer FRET efficiency based on the photobleaching rate of the donors present and the receptors deficiency. This method can be performed on most fluorescence microscopes and the photobleaching attenuation rate is usually independent of donor concentration.

• Lifetime measurements

We offer Fluorescence Lifetime Imaging Microscopy (FLIM). It can determine FRET efficiency based on changes in donor fluorescence lifetime. It is also possible to map microscopic images and the spatial distribution of fluorescence within living cells.

• Sensitized emission

We measured FRET efficiency by measuring changes in receptor emission intensity. FRET changes are observed when the distance or relative orientation between donors and acceptors varies. This method is simple and universally applicable.

The donor-acceptor fluorescence molecules we provide

Advantages of FRET

• Assays are performed under normal physiological conditions in living cells and do not require broken cells.
• Susceptible, enabling studies at the single-cell level.
• Can be combined with various instruments and techniques, such as microscopy, chromatography, electrophoresis, lost cell techniques,etc.
• Multi-application compatible.
• Modular system (lasers and filters are optional).

CD BioSciences is a pioneering biological company at the forefront of plant protein research. We have cutting-edge fluorescence microscopy, fluorophore selection, and accurate data analysis to help clients research plant protein-protein interactions. Please contact us if you are interested in our services, we are committed to helping you gain a deeper understanding of protein dynamics and advancing plant biology research.

For research use only, not for clinical use.