Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Super-resolution Fluorescence Microscopy

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...

Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

You might also read

Related Articles

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

Sort by

Same author

Insights Into Remote Ischemic Conditioning miRNA Effects on Brain Endothelial Cells During Ischemia-Reperfusion.

Microcirculation (New York, N.Y. : 1994)·2026

Same author

High-fidelity single-frame computational super-resolution using signal-preserving denoising-enabled deconvolution.

Nature communications·2026

Same author

Exploring extracellular vesicle surface markers and microRNA regulation following remote ischemic conditioning in patients with stroke; a randomized-controlled pilot study.

BMC neuroscience·2026

Same author

Direct Profiling of Extracellular Vesicle Surface Markers in Plasma: A Proof-of-Concept Study.

Biology of the cell·2025

Same author

A highly photostable monomeric red fluorescent protein for dual-color 3D STED and time-lapse 3D SIM imaging.

Nature methods·2025

Same author

GLP-1 Receptor Agonists as Treatment of Nondiabetic Ischemic Stroke: A Systematic Review and Meta-Analysis.

Stroke·2025

Same journal

RETRACTED: Bakshi et al. Crocin Inhibits Angiogenesis and Metastasis in Colon Cancer via TNF-α/NF-kB/VEGF Pathways. <i>Cells</i> 2022, <i>11</i>, 1502.

Cells·2026

Same journal

Correction: Verde et al. Molecular Mechanisms of Protein Aggregation in ALS-FTD: Focus on TDP-43 and Cellular Protective Responses. <i>Cells</i> 2025, <i>14</i>, 680.

Cells·2026

Same journal

Inflammation in Cardiomyopathies: Cellular Mechanisms Across Cardiac Phenotype.

Cells·2026

Same journal

IL-4/IL-13-Driven Dysregulation of Epidermal Lipid Metabolism in Atopic Dermatitis: An Immunometabolic Link Between Type 2 Inflammation and Barrier Dysfunction.

Cells·2026

Same journal

Activity of DNA- and RNA-Guided Prokaryotic Argonautes in Human Mitochondria.

Cells·2026

Same journal

Placental Pathophysiology in Maternal Psychoactive Substance Use: Biological, Clinical, and Forensic Perspectives.

Cells·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Sep 27, 2025

Author Spotlight: Advancements in Correlative Light and Electron Microscopy with Fluorescent Protein Preservation

Author Spotlight: Advancements in Correlative Light and Electron Microscopy with Fluorescent Protein Preservation

Published on: January 12, 2024

Improved Fluorescent Proteins for Dual-Colour Post-Embedding CLEM.

Dingming Peng^1,2, Na Li^1,3,4,5, Wenting He¹

¹Key Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.

|April 12, 2022

Summary

This summary is machine-generated.

Researchers developed new fluorescent proteins (FPs) to improve correlative light and electron microscopy (CLEM). These probes enhance signal-to-background ratio in Epon-embedded samples, enabling clearer visualization of cellular structures.

Keywords:

RSFP dual-colour CLEM high SBR probe development

More Related Videos

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

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

Published on: December 11, 2021

Dual-color Correlative Light and Electron Microscopy for the Visualization of Interactions between Mitochondria and Lysosomes

Dual-color Correlative Light and Electron Microscopy for the Visualization of Interactions between Mitochondria and Lysosomes

Published on: September 27, 2024

Related Experiment Videos

Last Updated: Sep 27, 2025

Author Spotlight: Advancements in Correlative Light and Electron Microscopy with Fluorescent Protein Preservation

Author Spotlight: Advancements in Correlative Light and Electron Microscopy with Fluorescent Protein Preservation

Published on: January 12, 2024

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

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

Published on: December 11, 2021

Dual-color Correlative Light and Electron Microscopy for the Visualization of Interactions between Mitochondria and Lysosomes

Dual-color Correlative Light and Electron Microscopy for the Visualization of Interactions between Mitochondria and Lysosomes

Published on: September 27, 2024

Area of Science:

Cell Biology
Microscopy Techniques
Biochemistry

Background:

Correlative light and electron microscopy (CLEM) offers high-precision imaging but is limited by available fluorescent proteins (FPs) and low signal-to-background ratio (SBR).
Epon resin embedding, crucial for preserving cellular ultrastructure, often quenches FP fluorescence, further complicating CLEM applications.

Purpose of the Study:

To develop novel fluorescent proteins (FPs) with improved performance for post-embedding CLEM.
To enhance the signal-to-background ratio (SBR) and enable dual-color imaging in Epon-embedded samples.

Main Methods:

Development and characterization of a green photoswitchable FP (mEosEM-E) with high on/off contrast.
Identification of a bright red fluorescent protein (RFP) mScarlet-H with superior SBR in Epon resin.
Application of subtraction-based CLEM (sCLEM) to reduce resin autofluorescence.
Dual-color CLEM imaging of nucleolar protein organization.

Main Results:

mEosEM-E demonstrated high photoswitching contrast, effectively reducing background autofluorescence via sCLEM.
mScarlet-H exhibited enhanced brightness and SBR compared to previously reported RFPs in Epon resin.
Successful dual-color post-Epon-embedding CLEM imaging with high SBR and no cross-talk was achieved.
Analysis of EM sample preparation effects on FP fluorescence preservation was conducted.

Conclusions:

Novel FPs, mEosEM-E and mScarlet-H, significantly advance post-embedding CLEM capabilities.
These probes facilitate high-resolution imaging of cellular ultrastructure and protein organization.
The findings provide guidance for developing future fluorescent probes for advanced microscopy.