Three-point repositioning of axes: three-dimensional alignment procedure for electron microscope tomography using three markers

A description is given of a new procedure to align series of tilted graphs, made with an electron microscope, for computer tomographic purposes. The procedure uses the coordinates of three projected markers to calculate parameters needed for the reconstruction. To that end the procedure computes the direction of the tilt-axis, the translation and rotation parameters, the tilt-angle of every micrograph, and the spatial coordinates of the individual markers with their centre of gravity as origin of the coordinate system. A searching technique, based on cross-correlation, is described to locate accurately the micrographs markers.