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

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

2.0K
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...
2.0K

You might also read

Related Articles

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

Sort by
Same author

A single-cell atlas identifies oncogenic transcriptional programs and immune escape mechanisms in CTCL.

Blood·2026
Same author

Nanoscale Fluorescent Characterizations and Origin Tracing of Extracellular Vesicles through Standard Preparation Methods.

ACS nano·2026
Same author

CDK9 is a dependency in GATA-3 driven and MCL-1 independent T-cell Lymphomas.

Blood cancer journal·2025
Same author

Ketoboronate as a Minimal Covalent-Reversible Tag for Targeted Lysosomal Degradation of Extracellular and Membrane Proteins.

Journal of the American Chemical Society·2025
Same author

Transcriptional Analysis of Effusion-Based Lymphoma Supports a Post-Germinal Center Origin and Specific Inflammatory Signal Background.

Cancers·2025
Same author

Total Synthesis of (-)-Neocucurbol C Enabled by Pattern Recognition and MHAT Cyclization.

Journal of the American Chemical Society·2025

Related Experiment Video

Updated: May 13, 2025

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

8.4K

Unveiling cellular communications through rapid pan-membrane-protein labeling.

Hirushi Gunasekara1, Yu-Shiuan Cheng1, Vanessa Perez-Silos2

  • 1Department of Chemistry, College of Liberal Arts and Sciences, University of Illinois Chicago, Chicago, IL, 60607, USA.

Nature Communications
|April 15, 2025
PubMed
Summary

This study introduces a novel amine crosslinking method for live cell surface protein labeling, enabling high-density multiplexed imaging. This technique visualizes dynamic cell interactions and protein trafficking in various cell types and in vivo.

More Related Videos

High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy
15:13

High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy

Published on: July 25, 2014

11.3K
A Label-free Technique for the Spatio-temporal Imaging of Single Cell Secretions
09:09

A Label-free Technique for the Spatio-temporal Imaging of Single Cell Secretions

Published on: November 23, 2015

8.5K

Related Experiment Videos

Last Updated: May 13, 2025

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

8.4K
High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy
15:13

High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy

Published on: July 25, 2014

11.3K
A Label-free Technique for the Spatio-temporal Imaging of Single Cell Secretions
09:09

A Label-free Technique for the Spatio-temporal Imaging of Single Cell Secretions

Published on: November 23, 2015

8.5K

Area of Science:

  • Cell Biology
  • Biochemistry
  • Microscopy

Background:

  • Dynamic protein distribution at the plasma membrane is crucial for cell communication.
  • Existing live imaging methods lack the speed and density for multiplexed labeling across diverse cell types.

Purpose of the Study:

  • To develop a rapid, high-density labeling method for live mammalian cell surface proteins.
  • To enable multiplexed live imaging of membrane dynamics and intercellular interactions.

Main Methods:

  • N-hydroxysuccinimide (NHS)-ester-based amine crosslinking of fluorescent dyes to cell surface proteins.
  • Live cell imaging, including multiplexed and 3D superresolution microscopy.
  • In vivo studies using a T cell lymphoma mouse model.

Main Results:

  • Successful uniform labeling of live mammalian cell surface proteins.
  • Visualization of transient membrane protein accumulation at cell-cell contacts and migration patterns.
  • Revealed biogenesis of membrane tunneling nanotubes and intercellular transfer.
  • Observed caveolin 1-dependent endocytosis and membrane remodeling in response to stimulation.
  • Demonstrated stable in vivo labeling for visualizing intercellular transfer in splenocytes.

Conclusions:

  • NHS-ester labeling provides a versatile tool for high-density, multiplexed live imaging of cell surface proteins.
  • The method facilitates the study of dynamic membrane processes, cell-cell interactions, and intercellular communication in vitro and in vivo.