Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

Super-resolution Fluorescence Microscopy

Super-resolution Fluorescence Microscopy

Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

The fluorescence-activating and absorption-shifting tag (FAST), a versatile protein marker for live plant cell imaging.

The Plant journal : for cell and molecular biology·2026

Same author

Unequal mitochondrial segregation promotes asymmetric fates during neurogenesis.

Nature communications·2025

Same author

Design and evaluation of a tripartite chemogenetic fluorescent reporter for visualizing ternary protein complexes.

Nature communications·2025

Same author

A tunable and versatile chemogenetic near-infrared fluorescent reporter.

Nature communications·2025

Same author

Mechanical forces and ligand binding modulate <i>Pseudomonas aeruginosa</i> PilY1 mechanosensitive protein.

Life science alliance·2025

Same author

Simultaneous detection of membrane contact dynamics and associated Ca<sup>2+</sup> signals by reversible chemogenetic reporters.

Nature communications·2024

Search research articles

Related Experiment Video

Updated: Jan 10, 2026

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

Super-Resolution Live-Cell Mapping of Protein-Protein Interactions Using Chemogenetic Split Reporters and Stimulated

Stephanie Board¹, Arnaud Gautier^1,2

¹Sorbonne Université, École Normale Supérieure, Chimie Physique et Chimie du Vivant (CPCV), CNRS, Université PSL, 75005, Paris, France.

Chembiochem : a European Journal of Chemical Biology

|November 26, 2025

Summary

This study introduces splitFAST2, a chemogenetic reporter, combined with STED microscopy to visualize protein-protein interactions (PPI) in live cells with super-resolution. This breakthrough enables precise mapping of PPIs and cellular structures below the diffraction limit.

Keywords:

chemogenetic split fluorescent reporters protein–protein interactions stimulated emission depletion nanoscopy super‐resolution imaging

More Related Videos

High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy

High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy

Published on: July 25, 2014

Photoactivated Localization Microscopy with Bimolecular Fluorescence Complementation BiFC-PALM

Photoactivated Localization Microscopy with Bimolecular Fluorescence Complementation BiFC-PALM

Published on: December 22, 2015

Related Experiment Videos

Last Updated: Jan 10, 2026

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy

High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy

Published on: July 25, 2014

Photoactivated Localization Microscopy with Bimolecular Fluorescence Complementation BiFC-PALM

Photoactivated Localization Microscopy with Bimolecular Fluorescence Complementation BiFC-PALM

Published on: December 22, 2015

Area of Science:

Cellular Biology
Microscopy
Biochemistry

Background:

Mapping protein-protein interactions (PPI) in live cells is crucial for understanding cellular functions.
Current super-resolution microscopy techniques face challenges in visualizing PPIs across entire live cells at high spatial resolution.

Purpose of the Study:

To develop a novel method for sub-diffraction imaging of PPIs in live cells.
To enhance the spatial resolution of PPI visualization using a combination of chemogenetic reporters and microscopy.

Main Methods:

Utilized the chemogenetic split fluorescent reporter splitFAST2.
Combined splitFAST2 with stimulated emission depletion (STED) microscopy for super-resolution imaging.
Applied the technique to map subcellular localization of both inducible and constitutive protein interactions.

Main Results:

Achieved precise and unambiguous localization of PPIs in live cells below the diffraction limit.
Demonstrated the capability to map subcellular localization of various protein interactions.
Developed low-background fluorescent probes for sub-diffraction imaging of cellular structures like actin and microtubules.

Conclusions:

The splitFAST2 and STED microscopy combination offers a powerful tool for high-resolution PPI mapping in live cells.
This approach overcomes limitations of existing methods, enabling detailed visualization of cellular processes.
The developed probes are effective for imaging complex cellular architectures with unprecedented detail.