The block was trimmed with a razor blade, removed, and mounted onto an aluminum pin using conductive epoxy glue (ITW Chemtronics, Enschede, The Netherlands) as previously described (38 (link)). The glue was then hardened at 60°C overnight and the block sputter coated with 2 nm platinum using a Q150R S sputter coater (Quorum Tech, East Sussex, UK).
Correlative SBF-SEM data were collected using a 3View2XP (Gatan, Pleasanton, CA) attached to a Sigma VP scanning electron microscope (Zeiss, Cambridge) as previously described (38 (link)). The scanning electron microscope was operated at an accelerating voltage of 2 kV with high current mode active, using a 20 μm aperture and chamber pressure of ∼8 Pa. A per pixel dwell time of 2 μs was used with a slice thickness of 50 nm. Images were acquired at 8,192 × 8,192 pixels with 8.7 nm pixel resolution (horizontal frame width of 71 μm). The entire volume comprised 420–520 slices to give an ∼1,500 μm3 image volume as inFig. 4 .
Correlative SBF-SEM data were collected using a 3View2XP (Gatan, Pleasanton, CA) attached to a Sigma VP scanning electron microscope (Zeiss, Cambridge) as previously described (38 (link)). The scanning electron microscope was operated at an accelerating voltage of 2 kV with high current mode active, using a 20 μm aperture and chamber pressure of ∼8 Pa. A per pixel dwell time of 2 μs was used with a slice thickness of 50 nm. Images were acquired at 8,192 × 8,192 pixels with 8.7 nm pixel resolution (horizontal frame width of 71 μm). The entire volume comprised 420–520 slices to give an ∼1,500 μm3 image volume as in
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