Fig. 4

Scheme of ex vivo and in vivo techniques to detect hypoxia and immune cells. a Ex-vivo techniques could be useful to create topographic maps of hypoxic-induced physical and functional barriers in immune excluded tumor areas, providing mechanistic highlights into the organization of immune exclusion determinants. Bioinformatic and spatial transcriptomics could investigate hypoxic transcriptional profiles in correlation with ICR or TIS. Immunohistochemistry of exogenous compounds or HIF-induced proteins coupled to digital pathology of immune cells could provide functional and morphological insights. b In-vivo techniques to detect gradients of hypoxia and immune infiltration may be useful to monitor immunotherapy efficiency in hypoxic tumors (clinical setting). To detect hypoxia and immune cells in patients, non-invasive methodologies are preferred