Zen black 2 1 sp3

Bright-field images were obtained with the help of a Zeiss Axiovert 135 microscope (Carl Zeiss Microscopy) equipped with a standard color camera (Canon 650D, Canon, Ota, Japan) and a low magnification objective (Leitz LD A-Plan 20×/0.3, Wetzlar, Germany). Fluorescence stainings were imaged by structured illumination microscopy using an ApoTome.2 mounted onto an Axio Imager M.2 microscope using an Axiocam MRm camera (Carl Zeiss Microscopy) via a 20× objective (Plan-Apochromat 20×/0.8 DIC, Carl Zeiss Microscopy) or a 63× oil objective (63×/1.4 Oil DIC M27, Carl Zeiss Microscopy) with ZEN 3.0 blue edition software (2464 × 2056 pixel, binning 2X2). For the recording of actin- and ZO1-staining in AC-1M-88 spheroids and Ishikawa monolayers, a Zeiss LSM 710 DUO confocal microscope was used. For spheroid visualization, single spheroids were transferred in PBS into a glass-bottomed dish (14 mm diameter, thickness no. 1·5, MatTek, Ashland, MA, USA) and imaged with an LGK 7872 ML8 argon ion laser with appropriate emission bandpass filters via a 20× objective (20×/0.8 M27) using Zen black 2.1 SP3 software (all Carl Zeiss Microscopy). Ishikawa cells were visualized using the same setup but with a 40× objective (LD C-Apochromat 40×/1.1 W Korr M27, Carl Zeiss Microscopy). Image processing was performed with the open-source, image-processing program Fiji based on ImageJ [49 (link)].