Single-fluorophore biosensors for sensitive and multiplexed detection of signalling activities

Nat Cell Biol. 2018 Oct;20(10):1215-1225. doi: 10.1038/s41556-018-0200-6. Epub 2018 Sep 24.


Unravelling the dynamic molecular interplay behind complex physiological processes such as neuronal plasticity requires the ability to both detect minute changes in biochemical states in response to physiological signals and track multiple signalling activities simultaneously. Fluorescent protein-based biosensors have enabled the real-time monitoring of dynamic signalling processes within the native context of living cells, yet most commonly used biosensors exhibit poor sensitivity (for example, due to low dynamic range) and are limited to imaging signalling activities in isolation. Here, we address this challenge by developing a suite of excitation ratiometric kinase activity biosensors that offer the highest reported dynamic range and enable the detection of subtle changes in signalling activity that could not be reliably detected previously, as well as a suite of single-fluorophore biosensors that enable the simultaneous tracking of as many as six distinct signalling activities in single living cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biosensing Techniques / methods*
  • Cell Tracking / methods*
  • Fluorescent Dyes / chemistry*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Mice
  • Microscopy, Confocal
  • NIH 3T3 Cells
  • PC12 Cells
  • Rats
  • Reproducibility of Results
  • Signal Transduction*
  • Single-Cell Analysis / methods*


  • Fluorescent Dyes
  • Green Fluorescent Proteins