Metastatic microtissues were fixed in 4% paraformaldehyde (PFA; Sigma-Aldrich) for 40 min, after either 3 or 24 hours of coculture in the various conditions. SDC microscopy (X-1 Yokogawa with Borealis modification), fitted with a 10× Plan Apo objective and a red channel filter set (excitation 561 nm), was used to collect z-stacks of the coculture microtissues. Maximal intensity projections were made from z-stacks using 1 μm as the step size and 70 μm as the thickness (>2 microtissues per condition analyzed with >4 fields of view, for 365 cells analyzed per condition on average). Cancer cell volume and sphericity were obtained from Imaris imaging analysis software (version 9.1.0, Bitplane AG, Zurich, Switzerland) according to the algorithm of (16 (link)).
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Imaging and Analysis of Metastatic Microtissues
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Corresponding Organization : Australian Regenerative Medicine Institute
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Variable analysis
independent variables
- Duration of coculture (3 hours or 24 hours)
- Cancer cell volume
- Cancer cell sphericity
- Fixation of microtissues in 4% paraformaldehyde (PFA) for 40 minutes
- Imaging using SDC microscopy (X-1 Yokogawa with Borealis modification) with a 10× Plan Apo objective and a red channel filter set (excitation 561 nm)
- Image analysis using Imaris imaging software (version 9.1.0) according to the algorithm from reference 16
- No positive or negative controls were explicitly mentioned in the provided information.
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