Volume Scanning Electron Microscopy of Serial Thick Tissue Sections with Gas Cluster Milling

A microscopy system comprising:

• a beam milling system configured for generating a beam of particles directed at a sample to irradiate the sample and to mill away successive surface layers from the sample wherein the beam of particles is directed at the sample at an angle of greater than 10 degrees to the surface of the sample;
• a scanning electron microscope system configured for irradiating the successive surface layers of the sample with an electron beam and for imaging the successive surface layers of the sample in response to the irradiation of the surface layer;
• and
• a processor configured for generating a three dimensional image of the sample based on the imaging of the successive layers of the sample.