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

Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

19.9K
Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
19.9K
Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

12.1K
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...
12.1K

You might also read

Related Articles

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

Sort by
Same author

Disparate immunity proteins independently inactivate an antibacterial nuclease toxin.

The Journal of biological chemistry·2026
Same author

Eosinophil-epithelial interactions mediate protective intestinal remodeling during food allergy.

bioRxiv : the preprint server for biology·2026
Same author

Summarizing the 2024 immunotherapy manual of the Canadian society of allergy and clinical immunology.

Allergy, asthma, and clinical immunology : official journal of the Canadian Society of Allergy and Clinical Immunology·2026
Same author

Microbial metabolism of food allergens determines the severity of IgE-mediated anaphylaxis.

Cell host & microbe·2026
Same author

Reply.

The Journal of allergy and clinical immunology·2026
Same author

Geographic Disparities in Evidence Investigating the Use of Biologics in Chronic Rhinosinusitis.

Journal of otolaryngology - head & neck surgery = Le Journal d'oto-rhino-laryngologie et de chirurgie cervico-faciale·2026
Same journal

Case Report of Gouty Arthritis Complicated by Septic Arthritis.

Journal of visualized experiments : JoVE·2026
Same journal

Pioglitazone in Skin Fibrosis: Mechanistic Rationale and Therapeutic Potential of Pioglitazone for Scarring Dermatoses.

Journal of visualized experiments : JoVE·2026
Same journal

A Combined Posterior Stable Prosthesis and Medial Condylar Sliding Osteotomy Technique for Kashin-Beck Osteoarthritis.

Journal of visualized experiments : JoVE·2026
Same journal

Fluorescence-Guided Laparoscopic Regional Anatomical Subsegmental Liver Resection Combined with Cholecystectomy through the Laennec Approach.

Journal of visualized experiments : JoVE·2026
Same journal

Olive Oil-Based Lipid Emulsion Ameliorates Immune Checkpoint Inhibitor-Induced Myocarditis via Inhibition of the NF-κB/NLRP3/IL-1β Pathway.

Journal of visualized experiments : JoVE·2026
Same journal

"Opening The Orifices And Alleviating Throat Obstruction" Four-step Acupuncture Method Combined with Motor Imagery Therapy for Post-Stroke Dysphagia.

Journal of visualized experiments : JoVE·2026
See all related articles

Related Experiment Video

Updated: Jan 11, 2026

Imaging Amyloid Tissues Stained with Luminescent Conjugated Oligothiophenes by Hyperspectral Confocal Microscopy and Fluorescence Lifetime Imaging
10:04

Imaging Amyloid Tissues Stained with Luminescent Conjugated Oligothiophenes by Hyperspectral Confocal Microscopy and Fluorescence Lifetime Imaging

Published on: October 20, 2017

14.0K

High-plex Imaging using Spectral Confocal Microscopy to Minimize Non-specific Tissue Fluorescence.

Saven Denha1, Vitoria M Olyntho1, Jake Colautti1

  • 1McMaster Immunology Research Centre, Department of Medicine, Faculty of Health Sciences, McMaster University; Schroeder Allergy and Immunology Research Institute, Faculty of Health Sciences, McMaster University.

Journal of Visualized Experiments : Jove
|November 17, 2025
PubMed
Summary
This summary is machine-generated.

Spectral Iterative Bleaching Extends Multiplexity (IBEX) enhances highly multiplexed imaging by reducing background noise and autofluorescence. This optimized technique allows for detailed spatial proteomic analysis in challenging tissues.

More Related Videos

Imaging Subcellular Structures in the Living Zebrafish Embryo
11:19

Imaging Subcellular Structures in the Living Zebrafish Embryo

Published on: April 2, 2016

12.3K
Confocal Microscopy Reveals Cell Surface Receptor Aggregation Through Image Correlation Spectroscopy
06:51

Confocal Microscopy Reveals Cell Surface Receptor Aggregation Through Image Correlation Spectroscopy

Published on: August 2, 2018

7.5K

Related Experiment Videos

Last Updated: Jan 11, 2026

Imaging Amyloid Tissues Stained with Luminescent Conjugated Oligothiophenes by Hyperspectral Confocal Microscopy and Fluorescence Lifetime Imaging
10:04

Imaging Amyloid Tissues Stained with Luminescent Conjugated Oligothiophenes by Hyperspectral Confocal Microscopy and Fluorescence Lifetime Imaging

Published on: October 20, 2017

14.0K
Imaging Subcellular Structures in the Living Zebrafish Embryo
11:19

Imaging Subcellular Structures in the Living Zebrafish Embryo

Published on: April 2, 2016

12.3K
Confocal Microscopy Reveals Cell Surface Receptor Aggregation Through Image Correlation Spectroscopy
06:51

Confocal Microscopy Reveals Cell Surface Receptor Aggregation Through Image Correlation Spectroscopy

Published on: August 2, 2018

7.5K

Area of Science:

  • Biomedical Imaging
  • Proteomics
  • Immunohistochemistry

Background:

  • Multiplexed imaging is crucial for studying cell function and interactions within native tissue environments.
  • Distinguishing true marker expression from background noise is essential for accurate data interpretation in fluorescence microscopy.
  • Conventional multiplexed imaging techniques can be limited by tissue autofluorescence and non-specific staining.

Purpose of the Study:

  • To adapt the Iterative Bleaching Extends Multiplexity (IBEX) protocol for improved signal-to-background ratio and reduced autofluorescence.
  • To develop a robust workflow for high-dimensional, spatially resolved proteomic analysis in complex tissues.
  • To demonstrate the efficacy of Spectral IBEX in challenging tissue models like human nasal polyps.

Main Methods:

  • Integration of spectral confocal detection with computational unmixing.
  • Incorporation of heparin blocking to minimize charge-based off-target binding.
  • Cyclic immunolabeling and fluorophore inactivation (IBEX) protocol adapted for spectral imaging.

Main Results:

  • Significant improvement in signal-to-background ratio and suppression of tissue autofluorescence.
  • Minimized spectral bleed-through and reduced overall acquisition time compared to conventional methods.
  • Successful imaging of 26 markers in human nasal polyp tissue, capturing structural, immune, and cell state information.

Conclusions:

  • Spectral IBEX provides a robust and broadly applicable strategy for multiplexed imaging.
  • The optimized approach is particularly suited for tissues with high autofluorescence and non-specific staining.
  • This method generates high-dimensional, spatially resolved proteomic data essential for understanding complex tissue architecture and cellular niches.