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Electron tomography (ET) is a method for obtaining 3D-structural information using TEM. The images of a tilt series are acquired over a wide range of viewing angles, and then these 2D-projection images are aligned and back-projected to generate a 3D-reconstruction. In biological research, ET can be applied to study unique structures, which are in the context of a cell. ET is ideally suited to reveal cell organelles ultra structure in the 5-20 nm resolution range.

This movie (click image to open) shows an aligned sequence of serial tilted views of the 120 nm thick section from nuclear membrane of NRK cell (normal rat kidney fibroblast). Images were collected (TEM Tecnai F20) at one degree intervals over a range from -65° to +65° at magnification of 8989x. Gold particles (10 nm) were used as fiducial markers to align the images of the raw dataset (H. Vihinen).

Aligned sequence of serial tilted views of the 120 nm thick section from nuclear membrane of normal rat kidney fibroblast cells

This movie (click image to open) shows a serial section reconstruction from nuclear membrane of dividing NRK cell, composed of 40 slices (H. Vihinen).

A serial section reconstruction from nuclear membrane of dividing NRK cell

This movie (click image to open) shows the modeled ER of an interphase CHO-K1 cell. The membranes of rough endoplasmic reticulum are coloured as yellow and the nuclear envelope as red (Puhka et al., 2007).

A model of endoplasmic reticulum of an interphase CHO-K1 cell

Image alignment and reconstruction are done on a Linux workstation utilizing IMOD software package (The Boulder Laboratory for 3-Dimensional Electron Microscopy of Cells, University of Colorado). The segmentation, modeling and visualization are done with Amira (TGS Inc.).

In addition to IMOD software reconstructions and models can be made and visualized by B-soft, Animabob (provided by Ken Chin-Purcell, Minnesota Supercomputer Center Inc.) or XTV (part of COMET refinement program, Skoglund et al. 1996, J. Struct. Biol. 117:173-188) softwares.

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Navigation Introduction User Information EM Personnel Instruments Methods Plastic embedding HPF-FS Pre-embedding immunolabelling Immunolabelling of cryo-sections Immunolabelling of acrylic sections Cytochemical staining CLEM Electron tomography Negative staining Specimen preparation for SEM Element analysis CryoEM 3View Publications Seminars and Courses Jokitalo Group	Methods Electron tomography Electron tomography (ET) is a method for obtaining 3D-structural information using TEM. The images of a tilt series are acquired over a wide range of viewing angles, and then these 2D-projection images are aligned and back-projected to generate a 3D-reconstruction. In biological research, ET can be applied to study unique structures, which are in the context of a cell. ET is ideally suited to reveal cell organelles ultra structure in the 5-20 nm resolution range. This movie (click image to open) shows an aligned sequence of serial tilted views of the 120 nm thick section from nuclear membrane of NRK cell (normal rat kidney fibroblast). Images were collected (TEM Tecnai F20) at one degree intervals over a range from -65° to +65° at magnification of 8989x. Gold particles (10 nm) were used as fiducial markers to align the images of the raw dataset (H. Vihinen). Aligned sequence of serial tilted views of the 120 nm thick section from nuclear membrane of normal rat kidney fibroblast cells This movie (click image to open) shows a serial section reconstruction from nuclear membrane of dividing NRK cell, composed of 40 slices (H. Vihinen). A serial section reconstruction from nuclear membrane of dividing NRK cell This movie (click image to open) shows the modeled ER of an interphase CHO-K1 cell. The membranes of rough endoplasmic reticulum are coloured as yellow and the nuclear envelope as red (Puhka et al., 2007). A model of endoplasmic reticulum of an interphase CHO-K1 cell Image alignment and reconstruction are done on a Linux workstation utilizing IMOD software package (The Boulder Laboratory for 3-Dimensional Electron Microscopy of Cells, University of Colorado). The segmentation, modeling and visualization are done with Amira (TGS Inc.). In addition to IMOD software reconstructions and models can be made and visualized by B-soft, Animabob (provided by Ken Chin-Purcell, Minnesota Supercomputer Center Inc.) or XTV (part of COMET refinement program, Skoglund et al. 1996, J. Struct. Biol. 117:173-188) softwares.
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