Journal
JOURNAL OF STRUCTURAL BIOLOGY
Volume 156, Issue 3, Pages 461-468Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.jsb.2006.05.010
Keywords
cryo-microscopy; electron tomography; fiducial markers; mitochondrion; keratin intermediate filaments; lamellar body
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We describe a novel approach for the accurate alignment of images in electron tomography of vitreous cryo-sections. Quantum dots, suspended in organic solvents at cryo- temperatures, are applied directly onto the sections and are subsequently used as fiducial markers to align the tilt series. Data collection can be performed from different regions of the vitreous sections, even when the sections touch the grid only at a few places. We present high-resolution tomograms of some organelles in cryo-sections of human skin cells using this method. The average error in image alignment was about 1 nm and the resolution was estimated to be 5-7 nm. Thus, the use of section-attached quantum dots as fiducial markers in electron tomography of vitreous cryo-sections facilitates high-resolution in situ 3D imaging of organelles and macromolecular complexes in their native hydrated state. (c) 2006 Elsevier Inc. All rights reserved.
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