Related Experiment Video
Updated: Jun 16, 2026
08:27
Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer
Published on: October 1, 2016
Indicators based on fluorescence resonance energy transfer (FRET)
Cold Spring Harbor Protocols
|February 12, 2010
Summary
Fluorescence Resonance Energy Transfer (FRET) is a versatile spectroscopic technique used to image cellular functions. New FRET indicators are engineered to respond to various biochemical and physiological signals in living cells.
Area of Science:
- Biophysics
- Cell Biology
- Spectroscopy
Background:
- Fluorescence Resonance Energy Transfer (FRET) is a spectroscopic technique measuring proximity and orientation of chromophores.
- It is widely used in biochemistry and cell biology to determine molecular distances and orientations.
- Recent advancements involve engineering molecules to alter FRET in response to cellular signals.
Purpose of the Study:
- To summarize the photophysical principles of FRET.
- To review the types of FRET indicators used for optical imaging.
- To highlight FRET's versatility in designing cellular probes.
Main Methods:
- Exploitation of FRET for optical imaging of cellular functions.
- Engineering macromolecules and molecular pairs to modulate FRET.
- Utilizing emission ratioing for quantifiable FRET measurements.
Main Results:
- FRET indicators can monitor diverse cellular signals including membrane potential, cAMP, protease activity, Ca(2+), CaM, protein heterodimerization, phosphorylation, and gene expression.
- FRET is general, nondestructive, and easily imaged.
- Emission ratioing offers reliable quantification and is suitable for high-speed imaging.
Conclusions:
- FRET is a highly versatile spectroscopic readout for designing new cellular probes.
- Its ability to be imaged and quantified makes it ideal for studying cellular dynamics.
- FRET-based indicators are crucial for advancing our understanding of biochemical and physiological processes in living cells.
Related Concept Videos
01:19Protein Dynamics in Living Cells
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
01:05Total Internal Reflection Fluorescence Microscopy
Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.