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

14.7K
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...
14.7K
Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

21.5K
Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
21.5K

You might also read

Related Articles

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

Sort by
Same author

Fluorophore unmixing based on bleaching and recovery kinetics using MCR-ALS.

Talanta·2021
Same author

An introduction to optical super-resolution microscopy for the adventurous biologist.

Methods and applications in fluorescence·2018
Same author

Antiviral treatment of influenza A (H1N1-09) guided by molecular resistance testing in aplasia after allo-SCT.

Bone marrow transplantation·2011
Same author

Household transmissibility and other characteristics of seasonal oseltamivir-resistant influenza A(H1N1) viruses, Germany, 2007-8.

Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin·2010
Same author

Oseltamivir-resistant influenza A(H1N1) viruses detected in Europe during season 2007-8 had epidemiologic and clinical characteristics similar to co-circulating susceptible A(H1N1) viruses.

Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin·2009
Same author

Immunotherapy of B-cell malignancies with genetically engineered human CD8+ natural killer T cells.

Leukemia·2006

Related Experiment Video

Updated: Mar 1, 2026

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

9.1K

Live-cell monochromatic dual-label sub-diffraction microscopy by mt-pcSOFI.

S Duwé1, W Vandenberg, P Dedecker

  • 1Laboratory for NanoBiology, Department of Chemistry, KU Leuven, Celestijnenlaan 200G, 3001 Leuven, Belgium. peter.dedecker@hotmail.com.

Chemical Communications (Cambridge, England)
|June 1, 2017
PubMed
Summary
This summary is machine-generated.

We developed multi-tau photochromic super-resolution optical fluctuation imaging (mt-pcSOFI) to image two spectrally identical fluorescent proteins simultaneously in living cells. This advanced technique uses blinking kinetics for unmixing, enabling dual-channel sub-diffraction microscopy.

More Related Videos

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
10:20

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Published on: September 5, 2019

8.8K
Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
14:12

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells

Published on: December 11, 2021

6.1K

Related Experiment Videos

Last Updated: Mar 1, 2026

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

9.1K
Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
10:20

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Published on: September 5, 2019

8.8K
Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
14:12

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells

Published on: December 11, 2021

6.1K

Area of Science:

  • Microscopy
  • Biophysics
  • Cell Biology

Background:

  • Super-resolution microscopy techniques are crucial for visualizing cellular structures.
  • Photochromic super-resolution optical fluctuation imaging (pcSOFI) offers high spatial resolution.
  • Distinguishing spectrally similar fluorescent probes remains a challenge in live-cell imaging.

Purpose of the Study:

  • To expand pcSOFI for monochromatic dual-channel sub-diffraction microscopy.
  • To develop a method for unmixing spectrally identical reversibly switchable fluorescent proteins (RSFPs).
  • To enable simultaneous imaging of two distinct structures in living cells using RSFPs.

Main Methods:

  • Development and application of multi-tau pcSOFI (mt-pcSOFI).
  • Utilizing blinking kinetics of RSFPs for spectral unmixing.
  • Imaging experiments in living cells using existing and novel RSFPs (ffDronpa-F).

Main Results:

  • Successfully demonstrated mt-pcSOFI for monochromatic dual-channel imaging.
  • Achieved unmixing of spectrally identical RSFPs based on their distinct blinking kinetics.
  • Showcased simultaneous imaging of two cellular structures using mt-pcSOFI and ffDronpa-F.

Conclusions:

  • mt-pcSOFI is a powerful extension of pcSOFI for dual-channel super-resolution imaging.
  • The method allows for simultaneous visualization of multiple targets labeled with spectrally similar RSFPs.
  • This technique advances live-cell imaging capabilities by overcoming spectral overlap limitations.