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Microbial Biosensors

Microbial biosensors are analytical devices that utilize living microbes to detect specific substances through measurable signals. These devices consist of two main components: biosensing organisms and signal-transducing elements. Biosensing organisms, such as Escherichia coli or Saccharomyces cerevisiae, are typically housed in multiwell plates connected to transducers, enabling rapid, real-time detection of target analytes.Signal Generation MechanismWhen a target analyte—such as...

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Human Pseudoislet System for Synchronous Assessment of Fluorescent Biosensor Dynamics and Hormone Secretory Profiles
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Visualization of cGMP with cGi biosensors.

Martin Thunemann1, Natalie Fomin, Christian Krawutschke

  • 1Interfakultäres Institut für Biochemie, Universität Tübingen, Tübingen, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|May 28, 2013
PubMed
Summary

New fluorescent biosensors allow visualization of cyclic guanosine 3'-5'-monophosphate (cGMP) dynamics in living cells. These tools help understand cGMP signaling, crucial for various physiological and pathological processes.

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Measurement of Cyclic Guanosine Monophosphate (cGMP) in Solid Tissues using Competitive Enzyme-Linked Immunosorbent Assay (ELISA)
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Measurement of Cyclic Guanosine Monophosphate (cGMP) in Solid Tissues using Competitive Enzyme-Linked Immunosorbent Assay (ELISA)
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Measurement of Cyclic Guanosine Monophosphate (cGMP) in Solid Tissues using Competitive Enzyme-Linked Immunosorbent Assay (ELISA)

Published on: July 3, 2025

Area of Science:

  • Cellular and Molecular Biology
  • Biochemistry
  • Physiology

Background:

  • Cyclic guanosine 3 -5 -monophosphate (cGMP) is a key signaling molecule with roles in physiology and disease.
  • Understanding cGMP signaling requires monitoring its spatiotemporal dynamics and compartmentalization within cells.
  • The distinction between global and local cGMP pools influences cellular functional outcomes.

Purpose of the Study:

  • To provide an overview of genetically encoded fluorescent biosensors for cGMP.
  • To detail the application of fluorescence resonance energy transfer (FRET)-based cGi-type sensors for cGMP imaging.
  • To offer practical protocols for cGMP visualization in living cells, tissues, and organisms.

Main Methods:

  • Overview of available cGMP biosensors, with a focus on FRET-based cGi-type sensors.
  • Detailed protocols for primary cell isolation, adenoviral expression of sensors, and FRET microscopy setup.
  • In-cell sensor calibration, data evaluation, and discussion of limitations and pitfalls.

Main Results:

  • Demonstration of FRET microscopy for visualizing cGMP dynamics in various murine cell types (smooth muscle cells, cerebellar granule neurons).
  • Established protocols adaptable for different cell types and cGMP indicators.
  • Provided guidelines for cGMP imaging in living systems.

Conclusions:

  • Genetically encoded fluorescent biosensors, particularly FRET-based cGi sensors, are powerful tools for studying cGMP signaling.
  • The presented protocols facilitate the visualization of cGMP in diverse biological contexts.
  • This work advances the understanding of cGMP's role in cellular functions and potential therapeutic interventions.