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

Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

11.2K
A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
11.2K

You might also read

Related Articles

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

Sort by
Same author

Clinical Pharmacy Services in Canadian Emergency Departments: A 2022 National Survey.

The Canadian journal of hospital pharmacy·2024
Same author

Epithelial Ovarian Cancer: Providing Evidence of Predisposition Genes.

International journal of environmental research and public health·2022
Same author

Non-Epithelial Ovarian Cancers: How Much Do We Really Know?

International journal of environmental research and public health·2022
Same author

Heterogeneity of Circulating Tumor Cell-Associated Genomic Gains in Breast Cancer and Its Association with the Host Immune Response.

Cancer research·2021
Same author

Three-dimensional reconstruction of the orbital retrobulbar vasculature.

Orbit (Amsterdam, Netherlands)·2021
Same author

Prkcz null mice show normal learning and memory.

Nature·2013

Related Experiment Video

Updated: May 5, 2026

A Rapid Method for Multispectral Fluorescence Imaging of Frozen Tissue Sections
07:50

A Rapid Method for Multispectral Fluorescence Imaging of Frozen Tissue Sections

Published on: March 30, 2020

8.2K

A Modified Bleaching Method for Multiplex Immunofluorescence Staining of FFPE Tissue Sections.

Dan Wang1, Alison Cheung1, Gordon E Mawdsley1

  • 1Biomarker Imaging Research Lab, Sunnybrook Research Institute.

Applied Immunohistochemistry & Molecular Morphology : AIMM
|October 7, 2024
PubMed
Summary

A modified photochemical bleaching method enhances multiplex immunofluorescence (mIF) staining by improving fluorophore removal and preserving tissue integrity. This technique reduces processing time and maintains antigenicity for biomarker co-localization studies.

More Related Videos

Author Spotlight: Multiplex Immunofluorescence Combined with Spatial Image Analysis for the Clinical and Biological Assessment of the Tumor Microenvironment
06:05

Author Spotlight: Multiplex Immunofluorescence Combined with Spatial Image Analysis for the Clinical and Biological Assessment of the Tumor Microenvironment

Published on: June 2, 2023

7.3K
DNA-barcode-based Multiplex Immunofluorescence Imaging to Analyze FFPE Specimens from Genetically Reprogrammed Murine Melanoma
09:52

DNA-barcode-based Multiplex Immunofluorescence Imaging to Analyze FFPE Specimens from Genetically Reprogrammed Murine Melanoma

Published on: June 6, 2025

76

Related Experiment Videos

Last Updated: May 5, 2026

A Rapid Method for Multispectral Fluorescence Imaging of Frozen Tissue Sections
07:50

A Rapid Method for Multispectral Fluorescence Imaging of Frozen Tissue Sections

Published on: March 30, 2020

8.2K
Author Spotlight: Multiplex Immunofluorescence Combined with Spatial Image Analysis for the Clinical and Biological Assessment of the Tumor Microenvironment
06:05

Author Spotlight: Multiplex Immunofluorescence Combined with Spatial Image Analysis for the Clinical and Biological Assessment of the Tumor Microenvironment

Published on: June 2, 2023

7.3K
DNA-barcode-based Multiplex Immunofluorescence Imaging to Analyze FFPE Specimens from Genetically Reprogrammed Murine Melanoma
09:52

DNA-barcode-based Multiplex Immunofluorescence Imaging to Analyze FFPE Specimens from Genetically Reprogrammed Murine Melanoma

Published on: June 6, 2025

76

Area of Science:

  • Biotechnology
  • Histology
  • Immunohistochemistry

Background:

  • Multiplex immunofluorescence (mIF) is crucial for biomarker profiling and analyzing biomarker co-relationships within tissue sections.
  • The standard Cell DIVE mIF platform uses alkaline hydrogen peroxide for sequential fluorophore inactivation, but faces challenges like suboptimal bleaching, tissue degradation, and loss of antigenicity.
  • These limitations hinder efficient and reliable mIF analysis, particularly in FFPE tissues.

Purpose of the Study:

  • To develop and evaluate a modified photochemical bleaching technique to overcome the limitations of standard mIF bleaching protocols.
  • To assess the efficiency of fluorophore removal, tissue integrity, and antigen immunogenicity using the modified method compared to the conventional approach.
  • To determine the impact of the modified technique on overall processing time and its utility for multiple rounds of staining and imaging.

Main Methods:

  • A modified photochemical bleaching method was developed, involving intense LED light exposure combined with hydrogen peroxide application.
  • This technique was applied in repeated staining/bleaching cycles on breast and other tissue sections.
  • Residual fluorescence signal, tissue integrity, and antigenicity were quantitatively and qualitatively assessed and compared against the conventional bleaching method.

Main Results:

  • The modified photochemical bleaching effectively removed fluorescence signals from previous staining rounds, yielding consistent results across multiple cycles.
  • Photochemical treatments using the modified method preserved sub-cellular structures and tissue antigenicity throughout the mIF process.
  • Processing time for an 8-round mIF procedure was reduced from 36 to 30 hours, with significantly less tissue degradation compared to the conventional method.

Conclusions:

  • The modified photochemical bleaching technique offers reliable fluorophore removal in mIF, significantly improving tissue integrity and antigen preservation.
  • This enhanced method reduces overall processing time, making it a valuable tool for studying biomarker co-localization in various tissues.
  • The technique facilitates more robust and efficient multiplex biomarker analysis, overcoming key limitations of conventional mIF protocols.