Characterisation of Mechanical Behaviour of an Oscillating Diamond Knife and Cut Surface Quality for Serial Block-face Scanning Electron Microscopy

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ConnectomX designs and manufactures the 'katana microtome,' a miniaturised ultramicrotome device used for Serial Block-Face Scanning Electron Microscopy (SBF-SEM). Installed inside a scanning electron microscope (SEM), the ultramicrotome uses a diamond knife to cut ultra-thin layers from biological samples. The SEM then images the newly exposed surface, allowing for the creation of high-resolution 3D biological structures, vital for understanding cellular functions and advancing disease treatments.The diamond knife oscillates side-by-side during cutting. This feature reduces cutting force and helps to microtome to achieve a consistent clean cuts at tens of nanometres. However, the exact mechanical behaviour of the oscillating knife, including in-plane oscillation and potential out-of-plane components remains uncertain.A comprehensive characterisation of the diamond knife movement and sample surface topology after each cut will help us confirm product specifications and performance, ensuring optimal image acquisition for 3D datasets. This information is crucial for optimising the diamond knife holder design to achieve even thinner cuts. Understanding and addressing this issue will provide a competitive edge over alternative SBF-SEM solutions, bolster our reputation, and expand our product's capabilities.