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

Super-resolution Fluorescence Microscopy01:37

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 developed.

You might also read

Related Articles

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

Sort by
Same author

Phenoxazines with a Phototransferable <i>N</i>-Acetyl Group and Acrylate Linker: Assembly by C-H Activation, Photoconversion to Fluorescent Dyes, Biolabeling, and Super-Resolution Imaging.

Journal of the American Chemical Society·2026
Same author

Synthesis of Benzo[<i>b</i>]thiophene 1,1-Dioxides via Pd-Catalyzed Sulfinylation of Aryl Triflates and Their Use as Large Stokes Shift Fluorophores for Multicolor Live-Cell Imaging with Self-Labeling Tags.

JACS Au·2026
Same author

Photoactivatable Carborhodol and Carborhodamine Dyes with One Cleavable Group: Synthesis, Spectra, and Fluorescence Nanoscopy Applications.

JACS Au·2026
Same author

Wavefront estimation through structured detection in laser scanning microscopy.

Biomedical optics express·2026
Same author

Correction of Background in Fluorescence Correlation Spectroscopy for Accurate Determination of Particle Number.

Biomolecules·2026
Same author

Fluorescent Diarylethenes With Polar Groups: Synthesis, Spectra, and Optical Microscopy Applications.

Chemistry (Weinheim an der Bergstrasse, Germany)·2025
Same journal

Denoising algorithm of Φ-OTDR systems based on adaptive fractional wavelet transform denoising.

Optics express·2026
Same journal

Millisecond photon-to-photon latency and high-speed volumetric projection system for optogenetics.

Optics express·2026
Same journal

Polarization-encoded coaxial structured light for high-precision 3D surface profilometry.

Optics express·2026
Same journal

Discrete freeform optical design based on collaborative optimization of point cloud and local normals.

Optics express·2026
Same journal

Ultrafast ghost imaging with 25 GHz speckle switching and wavelength-division multiplexing.

Optics express·2026
Same journal

Atomic vapor cells fabricated by femtosecond laser welding of standard-optical-quality glass.

Optics express·2026
See all related articles

Related Experiment Video

Updated: Jun 4, 2026

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy
12:51

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Simultaneous multi-lifetime multi-color STED imaging for colocalization analyses.

Johanna Bückers1, Dominik Wildanger, Giuseppe Vicidomini

  • 1Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany.

Optics Express
|March 4, 2011
PubMed
Summary
This summary is machine-generated.

We developed a STED microscope for multicolor colocalization. It uses fluorescence lifetime and wavelength separation, offering drift-insensitive analysis for up to three colors, ideal for precise biological imaging.

More Related Videos

Highly Multiplexed, Super-resolution Imaging of T Cells Using madSTORM
08:43

Highly Multiplexed, Super-resolution Imaging of T Cells Using madSTORM

Published on: June 24, 2017

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
11:06

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

Published on: June 30, 2018

Related Experiment Videos

Last Updated: Jun 4, 2026

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy
12:51

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Highly Multiplexed, Super-resolution Imaging of T Cells Using madSTORM
08:43

Highly Multiplexed, Super-resolution Imaging of T Cells Using madSTORM

Published on: June 24, 2017

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
11:06

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

Published on: June 30, 2018

Area of Science:

  • Microscopy
  • Biophysics
  • Spectroscopy

Background:

  • Colocalization experiments are crucial for understanding molecular interactions in cells.
  • Traditional multicolor microscopy techniques face challenges with spectral overlap and photobleaching.
  • Stimulated Emission Depletion (STED) microscopy offers super-resolution but requires optimization for multi-color analysis.

Purpose of the Study:

  • To present a novel STED microscope setup optimized for three-color colocalization.
  • To demonstrate a method for separating fluorescence signals using both lifetime and wavelength discrimination.
  • To provide a drift-insensitive approach for accurate colocalization analysis.

Main Methods:

  • Implementation of a STED microscope with optimized optical pathways for multi-color excitation and detection.
  • Utilizing fluorescence lifetime imaging microscopy (FLIM) to differentiate two fluorophores.
  • Employing wavelength discrimination for the third fluorescence channel.
  • Proposing an optional second STED beam for enhanced multicolor recording stability.

Main Results:

  • Successful separation of up to three distinct fluorescence labels in colocalization experiments.
  • Demonstrated drift-insensitivity of the lifetime-based separation method, enhancing colocalization accuracy.
  • Validated the system's suitability for precise analysis of molecular proximity and interactions.

Conclusions:

  • The developed STED microscope provides a robust and accurate platform for three-color colocalization studies.
  • The combined use of fluorescence lifetime and wavelength separation offers a versatile solution for complex biological imaging.
  • The proposed setup advances the capabilities of super-resolution microscopy for detailed cellular analysis.