The KUSA-A1 cells seeded in the dish holder on a 50-nm SiN film were stained with anti-mouse collagen antibody (1:50, Novus Biologicals USA, Colorado, USA) diluted in a 1:1 mixture of PBS and media (50 μl) for 30 min at 4°C, washed twice with the mixture solution, and stained with anti-rabbit IgG-conjugated 60-nm gold colloids (1:50, Cytodiagnostics Inc., Ontario, Canada) diluted in a 1:1 mixture of PBS and media (50 μl) for 30 min at 4°C. After washing twice, the holder was attached to the acrylic sample holder in the 50-nm SiN film and observed using the SE-ADM system.

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