Figure 3

Agarose microarray fabrication (with spheroids). (A) Microjetting bubble removal technique. (B) Spheroid gravitational transfer technique. (C) Water removal. (D) Agarose infiltration/mixing. (E) Final cassette position on PDMS negative mold. (F) 96 spheroids embedded in 12 × 8 agarose microarray attached to sectioning cassette from side view. Spheroids appear to be in the same sectioning plane (white dotted line) and parallel to the cassette base. As a frame of reference, a yellow dotted line shows spheroid offset in the z direction that would occur with a 2° misalignment of the agarose microarray to the cutting plane. (G) Images of a single block going through the sequential process starting with spheroid microarray pattered in water (step B above) through a stained paraffin section (large tissue fragments are placed in two wells instead of spheroids for easy identification of microarray orientation). Video available in Supplemental Video 1.