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ID: 194940.0, MPI für biophysikalische Chemie / Molekulare Biologie (Dr. Thomas M. Jovin)
Alignment of the cell nucleus from labeled proteins only for 4D in vivo imaging
Authors:Rieger, B.; Molenaar, C.; Dirks, R. W.; van Vliet, L. J.
Language:English
Date of Publication (YYYY-MM-DD):2004-08
Title of Journal:Microscopy Research and Technique
Volume:64
Start Page:142
End Page:150
Sequence Number of Article:doi:10.1002/jemt.20069
Copyright:C* 2004 by Wiley-Liss Inc.
Review Status:Peer-review
Audience:Not Specified
Abstract / Description:Studies of protein dynamics by 4D (3D + time) confocal microscopy in vivo are hampered by global cell motion. The time between the acquisitions of the 3D images is in the order of minutes. Therefore, it is not to be expected that the cell as a whole remains fixed in the water basin on the stage. This superimposes a motion on the protein dynamics that has to be removed. We present a robust registration technqiue to align the cell images that does not require the a priori establishment of point-to-point correspondences. Instead, it uses the distribution of the labeled proteins. After correction for the translation, the 3D rotation of the cell is estimated. A robust intrinsic body coordinate system is constructed via the inertia tensor from the instensity distribution. By combining basis transformation to this intrinsic coordinate system, we can calculated [sic! Stefan] the rotation matrix in a conceptual and comutational straightforward manner. We evaluated the performance of this approach in three experiments with human osteaoarcoma cells (U-2 OS), where the nuclear proteins Histon H4 and PML were visualized. The PML is concentrated in several dozen nuclear spots. Expression of Histon H4 results in a total nuclear staining. The registration results for both channels computed independently are very similar. Practically, this means that only the labeled material needs to be observed and still registration of the cell as a whole can be achieved.
Free Keywords:motion registration; confocal microscopy; time series
Last Change of the Resource (YYYY-MM-DD):2004-08-19
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für biophysikalische Chemie/Abt. Thomas Jovin / 060
External Affiliations:Qunatitative Imaging Group, Faculty of Applied Sciences, Delft University of Technology, Lorentzweg 1, 2628 CJ Delft, The Netherlands; Leiden University medical Center, Department of Molecular Cell Biology, Laboratory for Cytochemistry and Cytometry, Wssenaarseweg 72, 2300 RA Leiden, The Netherlands
Identifiers:URL:http://www3.interscience.wiley.com/cgi-bin/fulltex...
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