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Updated: Apr 12, 2026

FRET Microscopy for Real-time Monitoring of Signaling Events in Live Cells Using Unimolecular Biosensors
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Fluorescent protein biosensors applied to microphysiological systems.

Nina Senutovitch1, Lawrence Vernetti2, Robert Boltz1

  • 1University of Pittsburgh Drug Discovery Institute, Pittsburgh, PA 15260, USA University of Pittsburgh Department of Computational & Systems Biology, Pittsburgh, PA 15260, USA.

Experimental Biology and Medicine (Maywood, N.J.)
|May 21, 2015
PubMed
Summary

Fluorescent protein biosensors (FPBs) have evolved from earlier techniques to enable real-time monitoring of cellular events in microphysiological systems (MPSs). These advanced tools are crucial for understanding cell dynamics in both healthy and diseased states.

Keywords:
Microphysiology systemsfluorescence microscopyfluorescent probesfluorescent protein biosensorsfluorescent proteinshigh-content screening

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Area of Science:

  • Cell biology
  • Biotechnology
  • Microfluidics

Background:

  • Fluorescence microscopy has advanced significantly, enabling detailed analysis of cellular activities.
  • Fluorescent protein biosensors (FPBs) represent a major evolution in studying cellular dynamics.
  • Microphysiological systems (MPSs) offer a platform for advanced cellular research.

Purpose of the Study:

  • To review the evolution of fluorescence techniques for studying living cells.
  • To highlight the current role and applications of FPBs in MPSs.
  • To provide examples of FPBs in a liver MPS model.

Main Methods:

  • Evolution from fluorescent analog cytochemistry (FAC) to FPBs.
  • Development of advanced fluorescence microscopy techniques (e.g., super-resolution, high-content screening).
  • Engineering of fluorescent proteins for environmental sensitivity and targeted protein interaction.

Main Results:

  • FPBs allow real-time, spatiotemporal measurement of cellular events in MPSs.
  • FPBs enable monitoring of diverse physiological processes, including post-translational modifications and protein interactions.
  • Successful application of FPBs in a liver MPS model demonstrates their utility.

Conclusions:

  • FPBs are powerful tools for real-time cellular analysis in MPSs.
  • The development of FPBs has significantly enhanced our ability to study cellular dynamics.
  • FPBs hold great promise for advancing research in microphysiological systems.