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

Sequential changes in calcium transients during M phase regulate cardiomyocyte proliferation.

The Journal of cell biology·2026
Same author

Digitally enriching a screening population for pancreatic cancer using routine blood-based measures and clinical histories.

ArXiv·2026
Same author

Treatment of OSA using mandibular advancement versus CPAP in improving cardiovascular health.

The American journal of medicine·2026
Same author

Structural analyses uncover protease-adhesin interactions and c-di-GMP receptor regulation in sulfate-reducing bacteria.

Nature communications·2026
Same author

Robust antibiotic sensitization of pathogenic Pseudomonas aeruginosa via negative hysteresis in the cell envelope.

Nature communications·2026
Same author

RTP004 Peptide Binds to Botulinum Neurotoxin, Increases Cell Surface Binding, and Enhances Cellular SNAP-25 Cleavage.

Toxins·2026

Related Experiment Video

Updated: Jun 1, 2026

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications
13:14

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Published on: April 14, 2015

Circular permutation of red fluorescent proteins.

Bo Shui1, Qi Wang, Frank Lee

  • 1Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America.

Plos One
|June 8, 2011
PubMed
Summary

Researchers engineered circular fluorescent proteins (mCherry and mKate) for novel molecular sensors. Several variants retained high brightness, offering new tools for biological research and sensor development.

More Related Videos

Effect of Fluorescent Proteins on Fusion Partners Using Polyglutamine Toxicity Assays in Yeast
09:23

Effect of Fluorescent Proteins on Fusion Partners Using Polyglutamine Toxicity Assays in Yeast

Published on: November 28, 2018

Conducting Multiple Imaging Modes with One Fluorescence Microscope
08:32

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Related Experiment Videos

Last Updated: Jun 1, 2026

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications
13:14

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Published on: April 14, 2015

Effect of Fluorescent Proteins on Fusion Partners Using Polyglutamine Toxicity Assays in Yeast
09:23

Effect of Fluorescent Proteins on Fusion Partners Using Polyglutamine Toxicity Assays in Yeast

Published on: November 28, 2018

Conducting Multiple Imaging Modes with One Fluorescence Microscope
08:32

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • Circular permutation of fluorescent proteins is a key strategy for developing molecular sensors.
  • Engineering fluorescent proteins like mCherry and mKate can enhance their utility in various biological applications.

Purpose of the Study:

  • To systematically explore permutation sites in mCherry and mKate fluorescent proteins.
  • To identify circular permutants with high brightness and functional stability for sensor design.
  • To evaluate the structural and fluorescent properties of engineered variants.

Main Methods:

  • Tandem fusion template approach for generating circular permutants.
  • Quantitative fluorescence measurements to assess brightness and spectral properties.
  • Truncation analysis to determine essential terminal segments.
  • Structural evaluation of selected variants using biophysical techniques.

Main Results:

  • Identified circular permutants of mCherry and mKate with significant retention of parent brightness (>60% for mCherry, >90% for mKate).
  • Evaluated fluorescence contributions from neighboring mutations and identified essential N- and C-terminal segments.
  • Structural analysis of cp-mKate variants showed no major conformational changes and cis chromophore conformation.
  • Discovered four cp-mKate variants exhibiting over 80% of native fluorescence.

Conclusions:

  • Circularly permuted mCherry and mKate proteins are viable building blocks for molecular sensors.
  • Engineering strategies can yield highly fluorescent and structurally stable variants.
  • These novel fluorescent protein variants expand the toolkit for constructing advanced biosensors and for protein complementation assays.