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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.

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Related Experiment Video

Updated: Jun 26, 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

Photoactivatable mCherry for high-resolution two-color fluorescence microscopy.

Fedor V Subach1, George H Patterson, Suliana Manley

  • 1Department of Anatomy and Structural Biology, and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, New York 10461, USA.

Nature Methods
|January 27, 2009
PubMed
Summary

Researchers developed PAmCherry1, a red fluorescent protein that activates with violet light. This new protein offers improved performance for advanced microscopy, enabling detailed imaging of cellular structures.

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Assessing Protein Interactions in Live-Cells with FRET-Sensitized Emission
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Related Experiment Videos

Last Updated: Jun 26, 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

High-Accuracy Correction of 3D Chromatic Shifts in the Age of Super-Resolution Biological Imaging Using Chromagnon
08:18

High-Accuracy Correction of 3D Chromatic Shifts in the Age of Super-Resolution Biological Imaging Using Chromagnon

Published on: June 16, 2020

Assessing Protein Interactions in Live-Cells with FRET-Sensitized Emission
09:15

Assessing Protein Interactions in Live-Cells with FRET-Sensitized Emission

Published on: April 22, 2021

Area of Science:

  • Cell Biology
  • Biochemistry
  • Microscopy

Background:

  • Modern microscopy relies on photoactivatable fluorescent proteins for advanced imaging.
  • Existing red photoactivatable proteins have limitations in performance and application.
  • Development of improved photoactivatable probes is crucial for cellular research.

Purpose of the Study:

  • To develop novel mCherry variants that are initially dark and become red fluorescent upon violet-light irradiation.
  • To engineer a monomeric red photoactivatable protein with enhanced characteristics for microscopy applications.
  • To validate the utility of the developed protein in super-resolution imaging techniques.

Main Methods:

  • Selection of mCherry variants based on ensemble and single-molecule characteristics.
  • Characterization of PAmCherry1, including excitation/emission maxima (564/595 nm).
  • Performance comparison with existing monomeric red photoactivatable proteins.
  • Application in two-color photoactivated localization microscopy (PALM) imaging of cellular structures.

Main Results:

  • Developed PAmCherry1 with superior maturation speed, pH stability, photoactivation rate, contrast, and photostability.
  • Demonstrated PAmCherry1's suitability as a tag for two-color diffraction-limited and super-resolution imaging.
  • Successfully performed PALM imaging, resolving <200 nm clusters of transferrin receptor and clathrin light chain at <25 nm resolution.

Conclusions:

  • PAmCherry1 is a highly efficient monomeric red photoactivatable fluorescent protein.
  • Its enhanced properties make it an ideal probe for advanced intracellular imaging, including super-resolution microscopy.
  • PAmCherry1 facilitates detailed analysis of molecular interactions and cellular dynamics.