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

You might also read

Related Articles

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

Sort by

Same author

MAPK-driven glioma progression and reprogramming of the tumor-associated immune response.

Neuro-oncology·2026

Same author

APOE4 Drives Uniquely Dysfunctional Human Microglial States in Alzheimer's Disease.

bioRxiv : the preprint server for biology·2026

Same author

Tumor-draining lymph nodes in ovarian cancer lack germinal centers but harbor tumor-reactive memory B cells clonally linked to intra-tumoral B cells.

Immunity·2026

Same author

Spatial proteomics reveals CD8+ T cell signatures and cellular niches associated with active HIV-1 replication in lymph nodes.

bioRxiv : the preprint server for biology·2026

Same author

Data modeling the interplay between single-cell shape, single-cell protein expression, and tissue state.

Cell reports methods·2026

Same author

Glycosite mapping and in situ mass spectrometry imaging of MUC2 glycopeptides via on-slide mucinase digestion.

Nature communications·2026

Same journal

Isolation of Mesenchymal Stem Cell-Derived Extracellular Vesicles.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Modeling Melanoma Immune Surveillance by CAR-T Cells in Human Skin Organoids.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Stepwise Optimization of a Matrigel-Based In Vitro Angiogenesis Assay for Reproducible and Quantifiable 2D-Tube Formation Using HUVECs.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Quantifying Mechanical Properties of Fresh Ovarian Tissue with Optical Brillouin Microscopy.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

3D Chromatin Architecture During Early Development: New Methods and New Findings.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Metabolic Plasticity in Embryogenesis Throughout the Lens of NAD<sup></sup>.

Methods in molecular biology (Clifton, N.J.)·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Oct 13, 2025

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

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

Published on: April 7, 2023

High-Dimensional Tissue Profiling by Multiplexed Ion Beam Imaging.

Ofer Elhanani¹, Leeat Keren², Michael Angelo³

¹Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel.

Methods in Molecular Biology (Clifton, N.J.)

|November 12, 2021

Summary

This summary is machine-generated.

Multiplexed Ion Beam Imaging by Time of Flight (MIBI-TOF) provides high-dimensional, single-cell resolution imaging of clinical tissues. This method uses metal-labeled antibodies and mass spectrometry to visualize protein expression and localization in situ.

Keywords:

High-dimensional imaging Histology Imaging MIBI-TOF Mass spectrometry Multiplexed imaging Multiplexed ion beam imaging Pathology Proteomics Single cell

More Related Videos

Three-Dimensional Imaging of Tumor-Bearing Tissue Using the Iterative Bleaching Extends Multiplexity Approach

Three-Dimensional Imaging of Tumor-Bearing Tissue Using the Iterative Bleaching Extends Multiplexity Approach

Published on: April 25, 2025

High-Throughput Automated Multiplex Immunofluorescence Assays for Translational Research

High-Throughput Automated Multiplex Immunofluorescence Assays for Translational Research

Published on: June 10, 2025

Related Experiment Videos

Last Updated: Oct 13, 2025

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

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

Published on: April 7, 2023

Three-Dimensional Imaging of Tumor-Bearing Tissue Using the Iterative Bleaching Extends Multiplexity Approach

Three-Dimensional Imaging of Tumor-Bearing Tissue Using the Iterative Bleaching Extends Multiplexity Approach

Published on: April 25, 2025

High-Throughput Automated Multiplex Immunofluorescence Assays for Translational Research

High-Throughput Automated Multiplex Immunofluorescence Assays for Translational Research

Published on: June 10, 2025

Area of Science:

Biomedical Imaging
Mass Spectrometry
Proteomics

Background:

Multiplexed Ion Beam Imaging by Time of Flight (MIBI-TOF) is a powerful technique for in situ cellular analysis.
Existing methods may lack the resolution or dimensionality to fully characterize complex biological systems.

Purpose of the Study:

To detail the staining and imaging protocols for Multiplexed Ion Beam Imaging by Time of Flight (MIBI-TOF).
To enable researchers to implement MIBI-TOF for high-dimensional, single-cell analysis of clinical specimens.

Main Methods:

Tissue sections are stained with numerous metal-labeled antibodies.
Secondary ionization mass spectrometry is used to detect metal-antibody abundance and location.
This generates multi-dimensional images of protein expression and localization.

Main Results:

Successfully generated high-dimensional images of clinical specimens.
Achieved single-cell resolution for protein expression and localization.
Demonstrated the capability to visualize dozens of proteins simultaneously in situ.

Conclusions:

MIBI-TOF is a robust method for in situ proteomic profiling at single-cell resolution.
The described procedures facilitate the application of MIBI-TOF in clinical research.
This technique offers unprecedented insights into sub-cellular protein dynamics.