Abstract:
:Cryo-electron tomography of vitreous sections is currently the most promising technique for visualizing arbitrary regions of eukaryotic cells or tissue at molecular resolution. Despite significant progress in the sample preparation techniques over the past few years, the three dimensional reconstruction using electron tomography is not as simple as in plunge frozen samples for various reasons, but mainly due to the effects of irradiation on the sections and the resulting poor alignment. Here, we present a new algorithm, which can provide a useful three-dimensional marker model after investigation of hundreds to thousands of observations calculated using local cross-correlation throughout the tilt series. The observations are chosen according to their coherence to a particular model and assigned to virtual markers. Through this type of measurement a merit figure can be calculated, precisely estimating the quality of the reconstruction. The merit figures of this alignment method are comparable to those obtained with plunge frozen samples using fiducial gold markers. An additional advantage of the algorithm is the implicit detection of areas in the sections that behave as rigid bodies and can thus be properly reconstructed.
journal_name
J Struct Bioljournal_title
Journal of structural biologyauthors
Castaño-Díez D,Al-Amoudi A,Glynn AM,Seybert A,Frangakis ASdoi
10.1016/S1047-8477(08)00060-9subject
Has Abstractpub_date
2008-03-01 00:00:00pages
249-59issue
3eissn
1047-8477issn
1095-8657pii
S1047-8477(08)00060-9journal_volume
161pub_type
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