This high-resolution 3D tissue imaging video shows an intact mouse colorectal tumor stained to reveal immune organization across depth.

B220 is shown in green, CD3 in magenta, and nuclei in blue. Together, these markers map B and T cells in their native spatial context within the full tissue volume.

The workflow begins with wide-field screening of the entire cleared biopsy in 3D, enabling identification of tertiary lymphoid structure-rich regions dense in B220 signal. Regions of interest are then imaged at higher resolution, resolving cellular architecture while preserving the intact tissue block.

Because the sample remains structurally preserved, additional multiplex staining can be applied layer by layer, supporting deeper characterization of multiple TLSs within the same specimen.

This is volumetric, ground-truth 3D tissue imaging designed to reveal immune contexture at tissue scale with single-cell resolution.