In vivo imaging and histochemistry are combined in the cryosection labelling and intravital microscopy technique.

Intravital microscopy has been used extensively to study dynamic processes in the context of a living animal; however, only a limited number of fluorescent probes and mouse models are available. By contrast, many dyes and antibodies exist for the immuno-labelling of fixed tissue. Here we report a method that combines the advantages of histochemistry and in vivo imaging by correlating cryosection labelling with corresponding intravital microscopy images (CLIM). Using CLIM, we find that the presence of CD3(+) T cells correlates with mammary tumour cell migration. When CD4(+) and CD8(+) T-cell subsets are depleted, reduced tumour cell migration is observed. From these data we conclude that CLIM is a powerful tool to correlate intravital microscopy data with cryosection labelling data.