Fig. 1.

The fabrication workflow for creating 3D TEM for studying RMS in vitro. (A) PDMS is poured onto a glass slide. The cured polymer is cut into sections which are each punched with a biopsy punch to create the molds. (B) Collagen hydrogel is mixed with cells and pipetted into each mold. The hydrogel filled molds are placed in 12 well plates and thermally crosslinked for 45 min at 37 °C before being submerged with cell media for culture. (C) After 12 h of culture in the 3D constructs, cytotoxic drugs are added as desired to the culture media. Following a 48 or 96 h exposure to the cytotoxic drugs, the cells are analyzed using brightfield microscopy, immunocytochemistry, or live/dead viability assays.