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. 2021 Apr 23;7(17):eabc1323. doi: 10.1126/sciadv.abc1323

Fig. 3. Capturing real-time microcancer spheroid formation with live cell imaging on an inverted microscope and molecular characterizations of spheroids in microwell array.

Fig. 3

(A) Time progression of formation of five microcancers (LN229 cells) on our platform, imaged on an automated inverted microscope culture setup every 3 hours. Scale bar, 300 μm. (B) 2D projected distance between peripheral cell clusters in well 2 over 24 hours. These regions are shown in white circles and labeled in (A). (C) Reduction (%) from time “0” in 2D projected distance between peripheral cell clusters in well 2 over 24 hours. (D) Average reduction (%) from time “0” in 2D projected distance between peripheral cell clusters for all three wells over 24 hours. (E to G) Immunofluorescence detection of N-cadherin (red), 4′,6-diamidino-2-phenylindole nuclear staining (blue), and human mitochondria staining (green) in microcancers formed with LN229 cells and PDX. Tile of 12-well culture of LN229 microcancers (top) and PDX microcancers (bottom) imaged directly on chip (scale bar, 100 μm) (E). High-resolution images of individual microcancers for LN229 and PDX (scale bar, 20 μm) (F). Confocal z-stack images of PDX microcancer 3 days after cell seeding. Distance between each z-stack slice is 10 μm (scale bar, 20 μm) (G).