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

Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

5.6K
Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
5.6K
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

Platinum-based phosphorescent lifetime probes for the visualisation of G-quadruplex DNA in cells.

Chemical science·2026
Same author

Visualisation of adjuvant penetration into plant waxes by fluorescence of Nile Red.

Journal of materials chemistry. B·2026
Same author

Affordable, cleanroom-free millifluidic production of targeted lipid nanocarriers <i>via</i> additive manufacturing.

Lab on a chip·2026
Same author

Assembling Lipid Membrane Scaffolds on Microgel-Based Artificial Cells through Vesicle Fusion onto the Hydrogel Network.

ACS nano·2026
Same author

Estrogen-Functionalized Ru(II) Polypyridyl Complexes Self-Assemble into Aggregates and Exhibit Selective Phototoxicity against Breast Cancer Cells.

Inorganic chemistry·2025
Same author

Optical Probes for Cellular Imaging of G-quadruplexes: Beyond Fluorescence Intensity Probes.

Angewandte Chemie (International ed. in English)·2025

Related Experiment Video

Updated: Jun 5, 2025

Visualizing Intracellular SNARE Trafficking by Fluorescence Lifetime Imaging Microscopy
08:55

Visualizing Intracellular SNARE Trafficking by Fluorescence Lifetime Imaging Microscopy

Published on: December 29, 2017

9.5K

Imaging G-Quadruplex Nucleic Acids in Live Cells Using Thioflavin T and Fluorescence Lifetime Imaging Microscopy.

Tigerlily Bradford1, Peter A Summers1, Aatikah Majid1

  • 1Molecular Sciences Research Hub, Department of Chemistry, Imperial College London, London W12 0BZ, U.K.

Analytical Chemistry
|December 11, 2024
PubMed
Summary

Commercially available dyes Thioflavin T (ThT) and Thiazole Orange (TO) can now visualize G-quadruplex (G4) DNA structures in live cells. This fluorescence lifetime imaging microscopy (FLIM) method offers a simpler alternative for studying these important biological structures.

More Related Videos

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
09:45

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Published on: February 9, 2012

25.2K
4D Imaging of Protein Aggregation in Live Cells
08:59

4D Imaging of Protein Aggregation in Live Cells

Published on: April 5, 2013

17.3K

Related Experiment Videos

Last Updated: Jun 5, 2025

Visualizing Intracellular SNARE Trafficking by Fluorescence Lifetime Imaging Microscopy
08:55

Visualizing Intracellular SNARE Trafficking by Fluorescence Lifetime Imaging Microscopy

Published on: December 29, 2017

9.5K
Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
09:45

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Published on: February 9, 2012

25.2K
4D Imaging of Protein Aggregation in Live Cells
08:59

4D Imaging of Protein Aggregation in Live Cells

Published on: April 5, 2013

17.3K

Area of Science:

  • Cell Biology
  • Biophysics
  • Molecular Biology

Background:

  • Guanine-rich oligonucleotides form G-quadruplex (G4) structures in vivo.
  • These noncanonical DNA structures are implicated in crucial biological processes.
  • Direct visualization of G4s in live cells is essential for understanding their function.

Purpose of the Study:

  • To explore the use of common fluorescent dyes for G4 visualization.
  • To adapt fluorescence lifetime imaging microscopy (FLIM) for G4 detection using accessible reagents.
  • To establish a quantitative method for imaging G4s in live cells.

Main Methods:

  • Utilized fluorescence lifetime imaging microscopy (FLIM).
  • Investigated the fluorescence lifetime of Thioflavin T (ThT) and Thiazole Orange (TO) bound to different DNA topologies.
  • Applied FLIM with ThT to visualize G4s in live U2OS cells under varying conditions.

Main Results:

  • The fluorescence lifetime of ThT and TO is sensitive to DNA topology, with G4s exhibiting distinct, long decay times.
  • Successfully visualized G4 structures in live U2OS cells using FLIM with ThT.
  • Demonstrated G4 detection upon manipulation of G4 levels via competitive binding or nuclease treatment.

Conclusions:

  • Commercially available dyes ThT and TO can serve as effective probes for G4 structures based on their fluorescence lifetime.
  • FLIM using ThT provides a robust and accessible method for quantitative G4 imaging in live cells.
  • This approach simplifies G4 visualization, facilitating further research into their biological roles.