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Reporter Genes02:11

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
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: Jul 14, 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

Red-to-Near-Infrared Fluorescence Resonance Energy Transfer Reporters for Dual Imaging Applications.

Van Thi-Hong Tran1, Frans Ek2, Hasan Yassine1

  • 1Department of Biochemistry and Developmental Biology, Faculty of Medicine, University of Helsinki, Helsinki, Finland.

Small Methods
|July 13, 2026
PubMed
Summary

Researchers developed a new red-to-near-infrared fluorescent protein pair for enhanced multiplexed imaging in living cells. This tool allows simultaneous visualization of multiple signaling pathways, advancing cell biology research.

Keywords:
FLIMFRETbiosensorscell signalingfluorescent proteinsmRuby2miRFP670nano3

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Related Experiment Videos

Last Updated: Jul 14, 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

Conducting Multiple Imaging Modes with One Fluorescence Microscope
08:32

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Highly-Multiplexed Tissue Imaging with Raman Dyes
07:18

Highly-Multiplexed Tissue Imaging with Raman Dyes

Published on: April 21, 2022

Area of Science:

  • Cellular biology
  • Molecular imaging
  • Biophysics

Background:

  • Fluorescence resonance energy transfer (FRET) biosensors are crucial for visualizing protein interactions and signaling dynamics in live cells.
  • Existing FRET pairs often have overlapping spectra, limiting the ability to image multiple pathways simultaneously.
  • A lack of well-characterized near-infrared (NIR) acceptors hinders robust dual-FRET measurements within single cells.

Purpose of the Study:

  • To characterize a novel red-to-NIR Förster Resonance Energy Transfer (FRET) pair, mRuby2 and miRFP670nano3.
  • To assess its suitability for multiplexed imaging and simultaneous FRET measurements in living cells.
  • To demonstrate its application in studying complex signaling networks.

Main Methods:

  • Characterization of the mRuby2/miRFP670nano3 FRET pair.
  • Fluorescence Lifetime Imaging Microscopy (FLIM) for FRET efficiency and spectral bleed-through assessment.
  • Structural modeling to understand FRET efficiency.
  • Simultaneous FLIM-FRET imaging of signaling pathways (Akt, S6K, Src, ROCK) in living cells and 3D invasion assays.

Main Results:

  • The mRuby2/miRFP670nano3 pair exhibits efficient FRET with minimal spectral bleed-through.
  • This pair is compatible with existing Förster Resonance Energy Transfer (FRET) reporters like CFP/YFP.
  • It enables multiplexed imaging without complex spectral unmixing, outperforming alternative strategies.
  • Simultaneous imaging revealed coordinated signaling dynamics in Akt/S6K and Src/ROCK pathways.

Conclusions:

  • The mRuby2/miRFP670nano3 red-to-NIR Förster Resonance Energy Transfer (FRET) pair is a valuable tool for multiplexed imaging.
  • It facilitates simultaneous, quantitative analysis of multiple signaling pathways in living cells.
  • This advancement enables deeper insights into complex cellular signaling networks and dynamics.