This exciting article just published in Chemical Science by Professor W.E. Moerner and colleagues describes a novel method of introducing fluorophores of single-molecule quality into live bacterial cells for super-resolution imaging studies.
Single-molecule fluorescence imaging works by converting a dark fluorogen into a bright emitter. The really interesting aspect of this current study is that this conversion is achieved enzymatically. They have synthesised a nitro-aryl fluorogen (dark fluorogen) which is converted by a nitroreductase enzyme into a push-pull red-emitting fluorophore (bright emitter).
This new method allows the neutral dye molecule to enter the cell in a fluorescently ‘deactivated’ state. The substrate is then photoactivated by reaction with the enzyme, producing the fluorescent products – which is bright and detectable on the single-molecule level. The concentration can also be controlled by the level of substrate uptake – providing the opportunity for a variety of novel labeling systems.
The authors also present a detailed characterization of the spectral and photophysical properties of the fluorescent product, as well as the enzymatic kinetics in vitro.
Read the full article:
Enzymatic Activation of Nitro-Aryl Fluorogens in Live Bacterial Cells for Enzymatic Turnover-Activated Localization Microscopy
Marissa K. Lee, Jarrod Williams, Robert Twieg, Jianghong Rao and W.E. Moerner
Chem. Sci., 2012, DOI: 10.1039/C2SC21074F
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