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

ConnectomX设计并制造了“武士刀切片机”，这是一种用于连续块面扫描电子显微镜（SBF-SEM）的超薄切片机设备。超薄切片机安装在扫描电子显微镜（SEM）内，使用金刚石刀从生物样品上切割超薄层。扫描电子显微镜随后对新暴露的表面进行成像，从而创建高分辨率的3D生物结构，这对于了解细胞功能和推进疾病治疗至关重要。这一功能降低了切割力，有助于切片机在数十纳米处实现一致的干净切割。然而，振动刀的确切机械行为（包括面内振动和潜在的面外分量）仍不确定。对每次切割后金刚石刀运动和样品表面拓扑结构的全面表征将有助于我们确认产品规格和性能，确保3D数据集的最佳图像采集。这些信息对于优化金刚石刀保持器设计以实现更薄的切割至关重要。了解和解决这个问题将提供一个竞争优势，替代SBF-SEM解决方案，提高我们的声誉，并扩大我们的产品的能力。

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.