Multiplexing Autoradiography

Autoradiography, the direct imaging of radioactive distribution in tissue sections, is a powerful technique that has several key advantages for the validation of PET radiotracers. Using autoradiography, we can localize radiotracer uptake to neighbours of cells, and when multiplexed with additional radiotracers, fluorescent probes, or in situ tissue analysis, autoradiography can help to characterize the mechanism of radiotracer uptake and assess functional heterogeneity in tissue. In this chapter, the author outlines the basic ex vivo autoradiography protocol and shows how it can be multiplexed using dual radionuclides 18F and 14C. They also highlight where autoradiography can be combined with other technologies to provide synergistic information for interrogating spatial biology.