PITTSBURGHResearchers from Carnegie Mellon University's Molecular Biosensor and Imaging Center (MBIC) are turning up the brightness on a group of fluorescent probes called fluoromodules that are used to monitor biological activities of individual proteins in real-time. This latest advance enhances their fluormodule technology by causing it to glow an order of magnitude brighter than typical fluorescent proteins. The new fluoromodules are five- to seven-times brighter than enhanced green fluorescent protein (EGFP), a development that will open new avenues for research.
In a paper published online in the Journal of the American Chemical Society, MBIC researchers unveil a new class of dendron-based fluorogenic dyes called "dyedrons," that amplify the signal emitted by their fluoromodules.
"By using concepts borrowed from chemistry, the same concepts used in things like quantum dots and light harvesting solar cells, we were able to create a structure that acts like an antenna, intensifying the fluorescence of the entire fluoromodule," said Marcel Bruchez, associate research professor of chemistry and MBIC program director.
MBIC's fluoromodules are made up of a dye called a fluorogen and a fluorgen-activating protein (FAP). The FAP is genetically expressed in a cell and linked to a protein of interest, where it remains dark until it comes into contact with its associated fluorogen. When the protein and dye bind, the complex emits a fluorescent glow, allowing researchers to easily track the protein on the cell surface and within living cells. Fluoromodules are unique in that they do not need to be washed off for specific labeling, they come in a spectrum of colors, and they are more photostable than other fluorescent proteins.
To make the fluoromodules brighter, the researchers amplified the signal of one of their existing probes. They took one of their standard fluorogens, malachite green, and coupled it with another dy
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Carnegie Mellon University