Simultaneous alignment of dual-axis tilt series.
We present a strategy for the alignment of dual-axis tomographic series, based on reference points and simultaneous alignment of both series. Each series is first aligned individually, an affine transformation is determined to bring the two series in a unique reference system, and all experimental coordinates are combined in a single system of equations. In case of severe shrinkage, a global and a local refinement of the orientation parameters are performed to correct all minors misalignments. The strategy is illustrated on tomographic experiments performed on sections from plastic-embedded biological samples. The efficiency in correcting the misalignment of gold particles and in improving the quality of the reconstruction is documented both visually and quantitatively. In our approach every region of the tomogram is associated with its own orientation parameters and can be eventually reconstructed with the preferred algorithm. This is convenient in the computation of 3D averages of equivalent structures. A simulation experiment is presented to show that the performances of this approach are superior to those of the method of rotation in direct space.
Cantele
Francesca
F
Dipartimento di Chimica Strutturale, Università degli Studi di Milano, Italy.
Paccagnini
Eugenio
E
Pigino
Gaia
G
Lupetti
Pietro
P
Lanzavecchia
Salvatore
S
D002696:Chlamydomonas / Q000648:ultrastructure; D003198:Computer Simulation; D055032:Electron Microscope Tomography / Q000379:methods*; D005407:Flagella / Q000648:ultrastructure; D021621:Imaging, Three-Dimensional; D008854:Microscopy, Electron / Q000379:methods; D008962:Models, Theoretical