C11440 22c orca flash 4.0 v2

All high-resolution SRµCT acquisitions were performed using the white/pink beam setup of the SYRMEP (SYnchrotron Radiation for MEdical Physics) beamline of the Italian synchrotron ELETTRA (Trieste, Italy).
Whole paraffin embedded samples were scanned with a sample to detector distance of 500 mm. A a 16-bit, water-cooled sCMOS camera (Hamamatsu C11440-22C ORCA-Flash 4.0 v2) was used to acquire 3600 angular distributed projections with an exposure time of 50 ms and an isotropic voxel size of 2.33 µm. Scans were performed in a 360° offset regime, resulting in a scan time of 180 s. A 0.5 mm Si filter was used, resulting in a mean beam energy of 19.6 keV. All SRµCT data sets were reconstructed using SYRMEP Tomo Project^{48 (link)} software (STP Version 1.3.2). A single distance phase retrieval algorithm developed by Paganin et al.^{49 (link)} was used with a delta over beta ratio of 100. To image the entire tumours, 3–4 single acquisitions were performed, and the resulting reconstructions were manually stitched together using Fiji (NIH).

The X-ray phase-contrast CT scans were performed at the SYRMEP beamline of the Elettra Synchrotron^{72 (link)}, Trieste, Italy. All samples were scanned using the 2 GeV storage ring mode and a white (polychromatic) X-ray beam with the mean energy 25 keV, in combination with a sample-detector distance of 10 cm and exposure time per projection of 100 ms. The X-ray spectrum of the beam was filtered with 0.025 mm of molybdenum to reduce the contribution of low energy X-rays.
Projections were recorded with a 16-bit, air-cooled sCMOS camera (Hamamatsu C11440-22C ORCA-Flash 4.0 v2) (see^{73 (link)–76 (link)}). A 2.2 μm effective pixel size was set, using the variable optical zoom of the detecting system, corresponding to a field of view (FOV) of about 4.5 mm × 4.5 mm.
Using polychromatic radiation allows higher photon flux and thus decreases the scan duration of micro-CT experiments. For each sample in stages 28 to 42, a scan containing 900 projections was collected with a 0.2° angular step over 180° and total scanning time of around 2 min. Larger samples (stages 43 and 44) were scanned off-center over 360° (1800 projections) which allowed almost doubling of the FOV resulting in total scanning time of around 4 min.