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FIGURE 2 Summary of an analytic pipeline allowing the fine comparison of histochemical stain morphology and imaging mass cytometry <t>(IMC)</t> results within a single tissue section: example of an appendiceal sample. 1: The tissue section is produced based on a neutral formalin-fixed paraffin-embedded tissue sample and Alcian blue (AB) stained; 2: the AB-stained slide is digitalized; 3: the digital AB slide is used for the selection of the region of interest (ROI) for IMC analysis based on microscopic examination; 4: the ROI is positioned on the verso side of the AB-stained slide, which is used for IMC processing; 5: IMC acquisition of the ROI is performed; 6: the co-analysis of IMC data and digital AB slide allows the confrontation of morphological and immunophenotypic features until the single-cell level on the two images: note the exact overlapping and immunophenotyping between single cells within the Alcian blue (AB, CaseViewer, 3DHistech)) stain and IMC data (MCD Viewer, <t>Standard</t> <t>BioTools)</t> within the region of interest (ROI). (A) four neutrophile granulocytes with myeloperoxidase MPO positivity; (B) cytotoxic T cells CD8-positive within the appendiceal epithelium pan-keratin-positive; (C) three B cells CD20-positive and one neutrophile granulocyte MPO positive).
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FIGURE 2 Summary of an analytic pipeline allowing the fine comparison of histochemical stain morphology and imaging mass cytometry (IMC) results within a single tissue section: example of an appendiceal sample. 1: The tissue section is produced based on a neutral formalin-fixed paraffin-embedded tissue sample and Alcian blue (AB) stained; 2: the AB-stained slide is digitalized; 3: the digital AB slide is used for the selection of the region of interest (ROI) for IMC analysis based on microscopic examination; 4: the ROI is positioned on the verso side of the AB-stained slide, which is used for IMC processing; 5: IMC acquisition of the ROI is performed; 6: the co-analysis of IMC data and digital AB slide allows the confrontation of morphological and immunophenotypic features until the single-cell level on the two images: note the exact overlapping and immunophenotyping between single cells within the Alcian blue (AB, CaseViewer, 3DHistech)) stain and IMC data (MCD Viewer, Standard BioTools) within the region of interest (ROI). (A) four neutrophile granulocytes with myeloperoxidase MPO positivity; (B) cytotoxic T cells CD8-positive within the appendiceal epithelium pan-keratin-positive; (C) three B cells CD20-positive and one neutrophile granulocyte MPO positive).

Journal: Frontiers in immunology

Article Title: Coupling imaging mass cytometry with Alcian blue histochemical staining for a single-slide approach.

doi: 10.3389/fimmu.2024.1379154

Figure Lengend Snippet: FIGURE 2 Summary of an analytic pipeline allowing the fine comparison of histochemical stain morphology and imaging mass cytometry (IMC) results within a single tissue section: example of an appendiceal sample. 1: The tissue section is produced based on a neutral formalin-fixed paraffin-embedded tissue sample and Alcian blue (AB) stained; 2: the AB-stained slide is digitalized; 3: the digital AB slide is used for the selection of the region of interest (ROI) for IMC analysis based on microscopic examination; 4: the ROI is positioned on the verso side of the AB-stained slide, which is used for IMC processing; 5: IMC acquisition of the ROI is performed; 6: the co-analysis of IMC data and digital AB slide allows the confrontation of morphological and immunophenotypic features until the single-cell level on the two images: note the exact overlapping and immunophenotyping between single cells within the Alcian blue (AB, CaseViewer, 3DHistech)) stain and IMC data (MCD Viewer, Standard BioTools) within the region of interest (ROI). (A) four neutrophile granulocytes with myeloperoxidase MPO positivity; (B) cytotoxic T cells CD8-positive within the appendiceal epithelium pan-keratin-positive; (C) three B cells CD20-positive and one neutrophile granulocyte MPO positive).

Article Snippet: IMC analyses were conducted with the Hyperion® Imaging System technology (Standard BioTools) on tissue sections previously incubated (case #1) or not incubated (#2,#3,#4) with a pool of metal-coupled antibodies.

Techniques: Comparison, Staining, Imaging, Mass Cytometry, Produced, Selection