Journal: Frontiers in Oncology

Article Title: Intrinsic Differences in Spatiotemporal Organization and Stromal Cell Interactions Between Isogenic Lung Cancer Cells of Epithelial and Mesenchymal Phenotypes Revealed by High-Dimensional Single-Cell Analysis of Heterotypic 3D Spheroid Models

doi: 10.3389/fonc.2022.818437

Figure Lengend Snippet: List of IMC antibodies.

Article Snippet: 171 Yb , Histone H3 § , D1H2 , 201505 , Fluidigm , 1:4000.

Techniques:

Journal: Frontiers in Oncology

Article Title: Intrinsic Differences in Spatiotemporal Organization and Stromal Cell Interactions Between Isogenic Lung Cancer Cells of Epithelial and Mesenchymal Phenotypes Revealed by High-Dimensional Single-Cell Analysis of Heterotypic 3D Spheroid Models

doi: 10.3389/fonc.2022.818437

Figure Lengend Snippet: Characterization of spheroids using immunohistochemistry with fluorescent detection and imaging mass cytometry. HCC827-GFP monoculture, HCC827-GFP + SV80-dsRed co-culture, ER3-GFP + SV80-dsRed co-culture, and SV80-dsRed monoculture spheroids were stained with an imaging mass cytometry panel of 14 heavy metal-tagged antibodies ( Table 3 ). For each condition, five ROIs containing a single spheroid were ablated by a Hyperion imaging mass cytometer (Fluidigm, Inc.). Representative ROIs are displayed showing (A) GFP and RFP, (B) GFP, RFP and MKI67 (Ki67), (C) CDH1 (E-cadherin), and VIM (Vimentin), (D) GFP and EGFR, (E) and Collagen 1. DNA staining displayed in all images is a combination of Iridium intercalator stain (Ir191 and Ir193) and Histone H3. Images were pseudo-colored in MCD viewer software (Fluidigm) to enable visualization of multiple channels per ROI, and for each channel the minimum and maximum intensity display settings are manually set to be kept constant between the samples. Scalebar = 50 µm (F) Immunohistochemistry staining with the EGFRdel19 mutation-specific antibody and AF647 fluorescence tagged secondary antibody together with DAPI counterstain. Fluorescent images were taken with a Zeiss Collibri7 fluorescence microscope.

Article Snippet: 171 Yb , Histone H3 § , D1H2 , 201505 , Fluidigm , 1:4000.

Techniques: Immunohistochemistry, Imaging, Mass Cytometry, Co-Culture Assay, Staining, Cytometry, Software, Mutagenesis, Fluorescence, Microscopy