Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

2.1K
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...
2.1K

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

Fusion of magnetic fusogenic liposomes with model membranes for nanoparticle delivery.

Colloids and surfaces. B, Biointerfaces·2026
Same author

Chemogenetic Modulation of Luciferase Emission Color for Imaging and Sensing.

ACS sensors·2026
Same author

Unequal mitochondrial segregation promotes asymmetric fates during neurogenesis.

Nature communications·2025
Same author

Super-Resolution Live-Cell Mapping of Protein-Protein Interactions Using Chemogenetic Split Reporters and Stimulated Emission Depletion Microscopy.

Chembiochem : a European journal of chemical biology·2025
Same author

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

Nature communications·2025

Related Experiment Video

Updated: Jun 23, 2025

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy
10:41

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy

Published on: June 7, 2019

8.5K

Multiplexed In Vivo Imaging with Fluorescence Lifetime-Modulating Tags.

Lina El Hajji1, France Lam2, Maria Avtodeeva1

  • 1Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Laboratoire des Biomolécules, LBM, Paris, 75005, France.

Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|June 20, 2024
PubMed
Summary

Researchers developed novel fluorescence-activating and absorption-shifting tags (FASTs) to modulate fluorescence lifetime. This enables highly multiplexed imaging of multiple targets in live cells and organisms using a single fluorescent probe.

Keywords:
bioimagingfluorescence lifetime imagingfluorescent probesmultiplexed imagingprotein engineering

More Related Videos

Multiplexing Focused Ultrasound Stimulation with Fluorescence Microscopy
08:39

Multiplexing Focused Ultrasound Stimulation with Fluorescence Microscopy

Published on: January 7, 2019

8.2K
In Vivo Two-Color 2-Photon Imaging of Genetically-Tagged Reporter Cells in the Skin
05:45

In Vivo Two-Color 2-Photon Imaging of Genetically-Tagged Reporter Cells in the Skin

Published on: July 11, 2019

7.4K

Related Experiment Videos

Last Updated: Jun 23, 2025

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy
10:41

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy

Published on: June 7, 2019

8.5K
Multiplexing Focused Ultrasound Stimulation with Fluorescence Microscopy
08:39

Multiplexing Focused Ultrasound Stimulation with Fluorescence Microscopy

Published on: January 7, 2019

8.2K
In Vivo Two-Color 2-Photon Imaging of Genetically-Tagged Reporter Cells in the Skin
05:45

In Vivo Two-Color 2-Photon Imaging of Genetically-Tagged Reporter Cells in the Skin

Published on: July 11, 2019

7.4K

Area of Science:

  • Biophysics
  • Microscopy
  • Molecular Biology

Background:

  • Fluorescence lifetime imaging microscopy (FLIM) offers advanced multiplexing capabilities by distinguishing probes based on fluorescence lifetime.
  • Existing methods face limitations in achieving high-level multiplexing, especially in live biological systems.

Purpose of the Study:

  • To introduce novel fluorescence-activating and absorption-shifting tags (FASTs) for modulating fluorescence lifetime.
  • To demonstrate the utility of FASTs for highly multiplexed imaging in live cells and organisms.

Main Methods:

  • Development of FASTs engineered to alter the local environment of embedded fluorogenic 4-hydroxybenzylidene rhodanine (HBR) derivatives.
  • Utilizing changes in fluorescence lifetime to distinguish spectrally identical probes.
  • Application of FASTs for multiplexed imaging in live cells and zebrafish larvae.

Main Results:

  • FAST protein sequence modifications significantly altered chromophore environment and fluorescence lifetime.
  • Enabled multiplexed imaging of up to three targets in a single spectral channel using one HBR derivative.
  • Achieved successful imaging of six targets in live cells by combining FLIM and spectral multiplexing.

Conclusions:

  • FASTs provide a powerful tool for multicolor fluorescence lifetime multiplexing.
  • This approach significantly enhances multiplexing capacity in live-cell and in vivo imaging.
  • Opens new avenues for advanced multicolor imaging applications in biological research.