Dual-modality imaging of immunofluorescence and imaging mass cytometry for whole-slide imaging and accurate segmentation

• DOI: 10.1016/j.crmeth.2023.100595
• Publication Date: 2023-09-22T14:28:40Z
• Authors: Eun Na Kim; Phyllis Zixuan Chen; Dario Bressan; Monika Tripathi; Ahmad Miremadi; Massimiliano di P...; Lisa M. Coussens; Gregory J. Hannon; Rebecca C. Fitzge...; Lizhe Zhuang; Young Hwan Chang
• Keywords: Esophageal Neoplasms; CP: Imaging; Science; Q; Fluorescent Antibody Technique; QD415-436; Adenocarcinoma; Biochemistry; Article; Barrett Esophagus; Humans; TP248.13-248.65; Biotechnology; Image Cytometry

Imaging mass cytometry (IMC) is a powerful technique capable of detecting over 30 markers on a single slide. It has been increasingly used for single-cell-based spatial phenotyping in a wide range of samples. However, it only acquires a rectangle field of view (FOV) with a relatively small size and low image resolution, which hinders downstream analysis. Here, we reported a highly practical dual-modality imaging method that combines high-resolution immunofluorescence (IF) and high-dimensional IMC on the same tissue slide. Our computational pipeline uses the whole-slide image (WSI) of IF as a spatial reference and integrates small-FOV IMC into a WSI of IMC. The high-resolution IF images enable accurate single-cell segmentation to extract robust high-dimensional IMC features for downstream analysis. We applied this method in esophageal adenocarcinoma of different stages, identified the single-cell pathology landscape via reconstruction of WSI IMC images, and demonstrated the advantage of the dual-modality imaging strategy.