Related Experiment Video
Updated: Jun 26, 2025
A Rapid Method for Multispectral Fluorescence Imaging of Frozen Tissue Sections
Published on: March 30, 2020
Coupling imaging mass cytometry with Alcian blue histochemical staining for a single-slide approach
Patrice Hemon1, Danivanh Ben-Guigui1, Margaux Geier1,2
1Lymphocytes B, Autoimmunité et Immunothérapies (LBAI), Univ Brest, Inserm, Centre Hospitalier Universitaire (CHU) de Brest, UMR1227, Brest, France.
Insights
This study introduces a method combining histopathology with imaging mass cytometry (IMC) for better region selection and single-cell data analysis. This approach enhances the efficiency and depth of IMC tissue analysis.
Area of Science:
- Biomedical Imaging
- Cellular Biology
- Mass Spectrometry
Background:
- Imaging mass cytometry (IMC) enables multiplex immunophenotyping of cells in tissues.
- Current IMC limitations include 1-µm resolution and high analysis costs, restricting detailed histopathology and limiting analysis to small regions of interest (ROI).
Purpose of the Study:
- To develop a method for coupling histopathological analysis with IMC on a single tissue section.
- To improve ROI selection, reduce analysis time, and enable co-analysis of histopathological and immunophenotypic data at the single-cell level.
Main Methods:
- Developed a workflow integrating Alcian blue staining and digital imaging prior to destructive IMC analysis.
- Applied IMC to previously stained and digitalized tissue sections.
Main Results:
- Demonstrated the feasibility of performing IMC on Alcian blue-stained and digitalized tissue sections.
- The integrated method allows for improved ROI selection for IMC analysis.
Conclusions:
- Coupling histopathology with IMC on single tissue sections is feasible.
- This integrated approach enhances IMC's utility by improving ROI selection and enabling detailed co-analysis of histopathological and immunophenotypic data.
Abstract:
Imaging mass cytometry (IMC) is a metal mass spectrometry-based method allowing highly multiplex immunophenotyping of cells within tissue samples. However, some limitations of IMC are its 1-µm resolution and its time and costs of analysis limiting respectively the detailed histopathological analysis of IMC-produced images and its application to small selected tissue regions of interest (ROI) of one to few square millimeters. Coupling on a single-tissue section, IMC and histopathological analyses could permit a better selection of the ROI for IMC analysis as well as co-analysis of immunophenotyping and histopathological data until the single-cell level. The development of this method is the aim of the present study in which we point to the feasibility of applying the IMC process to tissue sections previously Alcian blue-stained and digitalized before IMC tissue destructive analyses. This method could help to improve the process of IMC in terms of ROI selection, time of analysis, and the confrontation between histopathological and immunophenotypic data of cells.
