Samples were incubated with secondary antibody for 72 h at 4°C and washed thoroughly in PBS before mounting onto slides with a 0.12 mm spacer (Grace Bin-Labs) in Tris-buffered glycerol-based mountant containing 0.5% p-phenylenediamine as an anti-fade.
Samples were washed three times with PBT, stained with 10 μg/ml Hoechst 33258 in PBT for 2 minutes, and washed three times with PBT before mounting onto slides with Fluoromount G. Samples were imaged on a Leica DMR fluorescence microscope using a Sensicam CCD camera and Slidebook software (Intelligent Imaging).
Cells were fixed in 4% paraformaldehyde in PBS for 20 min at room temperature, washed twice in PBS and permeabilized for 4 min in PBS containing 0.2% Triton X-100, and then blocked in TBS containing 2% BSA for 45 min. To visualize F-actin, cells were incubated with 1 µM TRITC-conjugated phalloidin (Sigma) and washed three times with PBS before mounting onto slides.
Coverslips were sputter coated with 4 nm chromium before mounting onto aluminium stubs (Agar Scientific) using Acheson ElectroDAG 1415M conducting adhesive (Agar Scientific) and sputter coating with 8 nm platinum-palladium 80/20.
Stain or paint shelf and allow it to dry before mounting onto the wall.
Teeth were wafered, decalcified, thin sectioned and stained, before being mounted onto a slide in order to facilitate counting of the growth layer groups [51].
Following further washing with PBS (2 × 1 ml) cells were counterstaining with 1 ml of PBS containing Hoechst 33258 0.6 μM, Invitrogen, UK) for 5 min, washed again with PBS (2 × 1 ml) before been mounted onto a glass microscope slide with Hydromount Vecta Mountant (approximately 50 μl, National Diagnostics, UK).
When the nitride-coated AAO template was mounted onto a steel plate before etching the nanotubes remained on the surface with the tips strongly intertwined.
After the samples had dried, each sample was mounted onto a microscope slide before a cover slip was added.
It carries a Atmega1284 microcontroller mounted onto a custom PCB designed by Bruce Land.
