Multiplex tissue imaging: An introduction to the scope and challenges

Abstract

Prompted by the multiplex study of Calvani et al (page 942), this editorial discusses opportunities and practicalities of a nextgeneration pathology, in which convergent technologies deep‐mine a tissue section to reveal unsuspected networks of spatial, social, and functional organization.

A robust multiplex immunofluorescence and digital pathology workflow for the characterisation of the tumour immune microenvironment

“…This can result in underexposure or oversaturation of the fluorophore when imaged with the multispectral scanning protocol, which has been optimised for a particular tissue thickness. Therefore, the use of tissue sections with regular thin cuts is highly recommended for mIF assays [7,31,32].…”

“…Natural variation in tissue thickness is expected and often seen in routine basic and special stains due to manual sectioning of the tissue. It is a recognised issue in manual IF assessment and one that is becoming increasingly apparent when developing artificial intelligence algorithms in histological images [31,32]. Compared to brightfield image analysis, assessment of mIF is uniquely sensitive to very subtle changes in tissue thickness, which can either increase or decrease the potential availability of exposed epitopes to fluorophore binding [33].…”

The Molecular Microscope (MMDXR) interpretation of thoracic and abdominal allograft biopsies: Putting things in perspective for the clinician

“…popularity and may render MMDx technology obsolete in the years to come. [20][21][22] The advent of three-dimensional intact-tissue sequencing of single-cell transcriptional states is particularly noteworthy. 23…”

“…Recent developments in histopathology, such as multiplex labeling, automated image analysis, and machine learning, are making it all the more important for gene expression data be analyzed in a way that surpasses the “blind grind and find” approach utilized by MMDx. Demand for platforms that merge spatial context with targeted mRNA and/or protein expression are growing in popularity and may render MMDx technology obsolete in the years to come 20‐22 . The advent of three‐dimensional intact‐tissue sequencing of single‐cell transcriptional states is particularly noteworthy 23 …”