Figure 1.

Tissue stitching and nuclear recognition. A–D: A representative colon section stained as described in the Methods with anti-CCR5 (detected by AlexaFluor350 and pseudo-colored in red), anti-CD68 (detected by AlexaFluor647 and pseudo-colored in white) and the nuclear stain Sytox Orange (pseudo-colored in blue) is depicted in its entirety. TissueFAXS software (TissueGnostics) stitched together this image of the complete colon section on the slide by aligning all overlapping 20x fields-of-view that had been sequentially acquired on the microscope (A). One 20x field-of-view is marked in A by the orange rectangle, which is then shown magnified in B–D. B: Three-color stain of cells in colon tissue, depicting CCR5 in red, CD68 in white and nuclei in blue. C: Nuclear stain Sytox Orange in monochrome grayscale. D: Recognition of the individual nuclei by the algorithm of the TissueQuest (TissueGnostics) software. Operator and software cell counts are indicated.