In FRET, two molecules that are fluorescent act as molecular beacons under the microscope, transferring energy between each other if they interact in the living cell, said Davidson, who directs the magnet labs Optical Microscopy program. With FRET, we can see that happen, but until now, we have only been able to monitor one biosensor at a time.
The new technique, called Dual FRET, is outlined in the paper Fluorescent Protein FRET Pairs for Ratiometric Imaging of Dual Biosensors. http://www.nature.com/nmeth/journal/v5/n5/abs/nmeth.1207.html
Further expanding the capabilities of optical microscopy, Davidson and his team worked with collaborators from the University of California, San Diego to create a new screening method for fluorescent proteins that makes them more stable under the microscope. These proteins are sensitive to light, which can bleach them out after a certain period of time. By making the proteins more stable, microscopists can observe live cell dynamics for longer periods of time. The paper describing their work, Improving the Photostability of Bright Monomeric Orange and Red Fluorescent Proteins, was published in the May 4 online edition of Nature Methods. http://www.nature.com/nmeth/journal/v4/n9/full/nmeth1083.html
Taken together, the new technique and tool are expected to speed up experiments and expand the utility of optical microscopy by allowing two dynamic processes inside a cell to be observed at once -- and for longer periods of time.
|Contact: Michael Davidson|
Florida State University