Over the past decade, our understanding of human diseases has rapidly grown from the rise of single-cell spatial biology. While conventional tissue imaging has focused on visualizing morphological features, the development of multiplex tissue imaging from fluorescence-based methods to DNA- and mass cytometry-based methods has allowed visualization of over 60 markers on a single tissue section. The advancement of spatial biology with a single-cell resolution has enabled the visualization of cell-cell interactions and the tissue microenvironment, a crucial part to understanding the mechanisms underlying pathogenesis. Alongside the development of extensive marker panels which can distinguish distinct cell phenotypes, multiplex tissue imaging has facilitated the analysis of high dimensional data to identify novel biomarkers and therapeutic targets, while considering the spatial context of the cellular environment. This mini-review provides an overview of the recent advancements in multiplex imaging technologies and examines how these methods have been used in exploring pathogenesis and biomarker discovery in cancer, autoimmune and infectious diseases. Keywords: Biomarker discovery; CNS, Central nervous system; CODEX, Co-detection by indexing; CTLA-4, Cytotoxic T-lymphocyte-associated protein 4; DSP, Digital Spatial Profiler; FF, Fresh-frozen; FFPE, Formalin-fixed paraffin-embedded; FOXP3, Forkhead box P3; HER2, Human epidermal growth factor receptor 2; HRP, Horseradish peroxidase; IDO, Indoleamine 2,3-dioxygenase; IHC, Immunohistochemistry; IL-10, Interleukin-10; IMC, Imaging Mass Cytometry; MCI, Mass Cytometry Imaging; MIBI, Multiplexed Ion Beam Imaging; MS, Multiple sclerosis; Multiplex tissue imaging; PD-L1, Programmed death-ligand 1; PD1, Programmed cell death protein 1; Pathogenesis; ROI, Regions of interest; SARS-CoV-2, Severe acute respiratory syndrome coronavirus 2; Spatial biology; Spatial proteomics; T1DM, Type 1 diabetes mellitus; TME, Tumor microenvironments; TNF-α, Tumor necrosis factor alpha; TOF, Time-of-flight; TSA, Tyramide signal amplification.