Abstract
In the evolving landscape of highly multiplexed imaging techniques that can be applied to study complex cellular microenvironments, this review characterizes the use of imaging mass cytometry (IMC) to study the human kidney. We provide technical details for antibody validation, cell segmentation, and data analysis specifically tailored to human kidney samples, and elaborate on phenotyping of kidney cell types and novel insights that IMC can provide regarding pathophysiological processes in the injured or diseased kidney. This review will provide the reader with the necessary background to understand both the power and the limitations of IMC and thus support better perception of how IMC analysis can improve our understanding of human disease pathogenesis and can be integrated with other technologies such as single cell sequencing and proteomics to provide spatial context to cellular data.
Highlights
The predominant methods for cell identification during the pathological analysis of formalinfixed paraffin-embedded (FFPE) samples from human renal core biopsy tissues include cell morphology and immunohistochemistry or immunofluorescence (IF)
Consistent with our previous study (Singh et al, 2019), 99.5 ± 4.6% of tubular cells were correctly identified by the Kidney-MAPPS pipeline compared with manual adjudication, indicating that identification of kidney cell populations is not impaired between reference and injured tissues
imaging mass cytometry (IMC) is a powerful platform for high-dimensional multiplexed parallel analysis of dozens of proteins localized in intact tissue, providing single-cell resolution while preserving the spatial relationship and morphological features
Summary
The predominant methods for cell identification during the pathological analysis of formalinfixed paraffin-embedded (FFPE) samples from human renal core biopsy tissues include cell morphology and immunohistochemistry or immunofluorescence (IF). The linearity of the mass spectrometry detection of the heavy metals conjugated to each antibody provides a quantitative assessment of the relative expression levels for the respective antigens, allowing researchers to detect changes not just in cell numbers and localization, and in cell differentiation and signaling pathway activation.
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