Dual-modality imaging of immunofluorescence and imaging mass cytometry for high-resolution whole slide imaging with accurate single-cell segmentation

Imaging mass cytometry (IMC) is a powerful multiplexed tissue imaging technology that allows simultaneous detection of more than 30 makers 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 small, rectangle field of view (FOV) with a low image resolution that hinders downstream analysis. Here, we reported a highly practical dual-modality imaging method that combines high-resolution immunofluorescence (IF) and high-dementional IMC on the same tissue slide. Our computational pipeline uses the whole slide image (WSI) of IF as spatial reference, 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.成像质谱流式细胞术（Imaging mass cytometry, IMC）是一种强大的多重组织成像技术，可在单张组织玻片上同时检测超过30种标志物。该技术已被越来越多地应用于多种样本的基于单细胞的空间表型分析中。然而，其仅能获取小尺寸矩形视野（FOV）的图像且分辨率较低，这一局限会对下游分析造成阻碍。本研究报道了一种实用性极强的双模态成像方法，可在同一张组织玻片上结合高分辨率免疫荧光（immunofluorescence, IF）与高维度成像质谱流式细胞术。我们的计算管线以免疫荧光的全玻片图像（WSI）作为空间参照，将小视野的成像质谱流式细胞术图像整合为成像质谱流式细胞术的全玻片图像。高分辨率的免疫荧光图像可实现精准的单细胞分割，从而提取出稳健的高维度成像质谱流式细胞术特征用于下游分析。我们将该方法应用于不同分期的食管腺癌中，通过重构成像质谱流式细胞术的全玻片图像解析出单细胞病理学图谱，并验证了该双模态成像策略的优势。