Imaging Mass Cytometry to Decipher the Maturation of Tertiary Lymphoid Structures

In the realm of inflammation, including conditions like cancers, infections, and autoimmune diseases, the emergence of tertiary lymphoid structures (TLS) holds significant implications for prognosis and treatment response. Yet, traditional methodologies such as immunohistochemistry and immunofluorescence falter in capturing the nuanced complexities of TLS, necessitating innovative approaches, like imaging mass cytometry (IMC), to unravel their significance. With the capacity to concurrently assess nearly 40 markers within the same tissue section, IMC transcends the constraints of traditional approaches.This chapter delineates the IMC methodology tailored for TLS visualization and subsequent quantification. By leveraging IMC on formalin-fixed, paraffin-embedded (FFPE) sections of human colorectal cancer (CRC) tissues, we aim to establish a standardized protocol that elucidates the intricacies of TLS dynamics and composition.