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

You might also read

Related Articles

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

Sort by
Same author

A disulfide-Fe<sup>3+</sup> crosslinking strategy for tough hydrogel networks with complete photo- and biochemical degradability.

Materials horizons·2026
Same author

Nanobody-Based Biotemplating Enables Nanoscale-Precision Metallization of Cellular, Tissue, and Plant Architectures.

ACS applied materials & interfaces·2026
Same author

Source-Free Domain-Adaptive Semi-Supervised Learning for Object Detection in CCTV Images.

Sensors (Basel, Switzerland)·2026
Same author

Multidirectional Trunk Movements Reveal Hidden Symmetry Loss in Stroke: An Electromyography-Based Comparative Study.

Medicina (Kaunas, Lithuania)·2025
Same author

Highly accurate image registration for 3D multiplexed cyclic imaging using dense labeling in expandable tissue gels.

PLoS biology·2025
Same author

The Effects of Upper Extremity Closed Kinetic Chain Exercise Combined with Biofeedback in Stroke Survivors: A Pilot Study.

NeuroRehabilitation·2025

Related Experiment Video

Updated: Jun 16, 2025

Multiplexed Barcoding Image Analysis for Immunoprofiling and Spatial Mapping Characterization in the Single-Cell Analysis of Paraffin Tissue Samples
08:18

Multiplexed Barcoding Image Analysis for Immunoprofiling and Spatial Mapping Characterization in the Single-Cell Analysis of Paraffin Tissue Samples

Published on: April 7, 2023

1.6K

Doubling multiplexed imaging capability via spatial expression pattern-guided protein pairing and computational

Gyuri Kim1, Hyejin Shin2, Minho Eom1

  • 1School of Electrical Engineering, KAIST, Daejeon, Republic of Korea.

Communications Biology
|June 14, 2025
PubMed
Summary

SEPARATE (Spatial Expression PAttern-guided paiRing And unmixing of proTEins) reduces 3D multiplexed imaging cycles by half. This novel method uses neural networks to unmix signals from two proteins labeled with the same fluorophore based on spatial patterns.

More Related Videos

Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging
08:40

Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging

Published on: April 8, 2016

12.8K
Spatial Profiling of Protein and RNA Expression in Tissue: An Approach to Fine-Tune Virtual Microdissection
09:19

Spatial Profiling of Protein and RNA Expression in Tissue: An Approach to Fine-Tune Virtual Microdissection

Published on: July 6, 2022

4.9K

Related Experiment Videos

Last Updated: Jun 16, 2025

Multiplexed Barcoding Image Analysis for Immunoprofiling and Spatial Mapping Characterization in the Single-Cell Analysis of Paraffin Tissue Samples
08:18

Multiplexed Barcoding Image Analysis for Immunoprofiling and Spatial Mapping Characterization in the Single-Cell Analysis of Paraffin Tissue Samples

Published on: April 7, 2023

1.6K
Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging
08:40

Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging

Published on: April 8, 2016

12.8K
Spatial Profiling of Protein and RNA Expression in Tissue: An Approach to Fine-Tune Virtual Microdissection
09:19

Spatial Profiling of Protein and RNA Expression in Tissue: An Approach to Fine-Tune Virtual Microdissection

Published on: July 6, 2022

4.9K

Area of Science:

  • Neuroscience
  • Microscopy
  • Biotechnology

Background:

  • Three-dimensional (3D) multiplexed fluorescence imaging is crucial for neuroscience research.
  • Cyclic immunofluorescence is a common 2D technique but is time-consuming for 3D applications due to long staining processes for thick specimens.

Purpose of the Study:

  • To develop a novel method, SEPARATE (Spatial Expression PAttern-guided paiRing And unmixing of proTEins), to significantly reduce the number of cycles required for 3D multiplexed imaging.
  • To enable efficient and faster volumetric imaging of multiple proteins in biological samples.

Main Methods:

  • SEPARATE utilizes a neural network to unmix signals from two proteins labeled with the same fluorophore.
  • A feature extraction network quantifies spatial expression patterns to determine protein pairs, generating feature-based distances.
  • Protein separation networks are then used for signal unmixing.

Main Results:

  • The feature extraction network demonstrated a high correlation between spatial pattern distinction and signal unmixing performance across ten proteins.
  • The method successfully reduced the number of imaging cycles by half.
  • Volumetric multiplexed imaging of six proteins using only three fluorophores was achieved.

Conclusions:

  • SEPARATE offers a significant advancement in 3D multiplexed fluorescence imaging by reducing experimental time and complexity.
  • The method's reliance on spatial expression patterns and neural network unmixing provides a robust approach for high-plex 3D imaging.
  • This technique has the potential to accelerate discoveries in neuroscience and other fields requiring detailed spatial protein analysis.