Reconstruction of single particles from

random conical tilt pairs using Pawel's methods

Date 7/30/97 Ramani

Updated 8/7/97 Doryen Bubeck

This procedure is to be used when there is no reference. In the Particle Selection phase of the procedure the spider formated micrographs are windowed in WEB. The tilt pairs corrdinates are selected and the corresponding partlices windowed. The following particles are Aligned using a reference free procedure and categorized through a heirarchical clustering. Then an Angular file is created and used in an iterative 3d reconstruction. In the end these reconstructions are merged.




Particle Selection

Important things to remember are:




Alignment




Creation of the ANGULAR FILE

and the 3D reconstruction.




Merged Reconstruction




Results

The final reconstruction could be used as a reference for future projection matching procedures provided it has no missing angular information. Even if the entire missing angular range is not covered in the merged reconstruction it will still present a better reconstruction as it atleast covers a part of the missing angular range compared to a reconstruction from just a single view. For obtaining reconstructions from reference projections click here.