The technique is particularly useful for 3D reconstructions of particles such as macromolecular complexes and viruses or for analysis of the organisation of subcellular compartments. Samples that have been chemically or cryo-fixed are prepared as semi-thin sections (less than 250 nm) or particle suspension and imaged in the TEM over a range of tilt angles (e.g. 0.5° increments through a 140° tilt series). During imaging, sophisticated TEM software ensures that sequential images are aligned, focused correctly and exposure optimised to facilitate subsequent image processing where the 2D series will be reconstructed into 3D images and models. More information about tomography can be found at the sites of other groups, including Boulder and Utrecht.
Our 3D electron tomography capabilities are now complemented with 3View 3D-EM imaging that takes serial block face images through a cell or tissue volume. This allows 1-100µm3 volumes of cells and tissues to be easiliy imaged and rendered.
Sample protocols:
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