Three-dimensional scanning transmission electron microscopy of biological specimens

Journal Article · Fri Jan 01 00:00:00 EST 2010 · Microscopy and Microanalysis

DOI:https://doi.org/10.1017/S1431927609991280· OSTI ID:972015

De Jonge, Niels ^[1]; Sougrat, Rachid ^[1]; Northan, Brian ^[2]; Pennycook, Stephen J ^[1]

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1. ORNL
2. Media Cybernetics, Inc.

A three-dimensional (3D) reconstruction of the cytoskeleton and a clathrin-coated pit in mammalian cells has been achieved from a focal-series of images recorded in an aberration-corrected scanning transmission electron microscope (STEM). The specimen was a metallic replica of the biological structure comprising Pt nanoparticles 2 - 3 nm in diameter, with a high stability under electron beam radiation. The 3D dataset was processed by an automated deconvolution procedure. The lateral resolution was 1.1 nm, set by pixel size. Particles differing by only 10 nm in vertical position were identified as separate objects with greater than 20% dip in contrast between them. We refer to this value as the axial resolution of the deconvolution or reconstruction, the ability to recognize two objects, which were unresolved in the original data set. The precision of the height determination was 0.2 nm. The resolution of the reconstruction is comparable to that achieved by tilt-series transmission electron microscopy (TEM). However, the focal-series method does not require mechanical tilting and is therefore much faster. 3D STEM images were also recorded of the Golgi ribbon in conventional thin sections containing 3T3 cells with a comparable axial resolution in the deconvolved data set.

• Research Organization: Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). High Temperature Materials Lab. (HTML)
• Sponsoring Organization: USDOE Laboratory Directed Research and Development (LDRD) Program; Work for Others (WFO)
• DOE Contract Number: DE-AC05-00OR22725
• OSTI ID: 972015
• Journal Information: Microscopy and Microanalysis, Vol. 16, Issue 1; ISSN 1431-9276
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
37 INORGANIC
ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE
BIOLOGICAL MATERIALS
SCANNING ELECTRON MICROSCOPY
TRANSMISSION ELECTRON MICROSCOPY
THREE-DIMENSIONAL CALCULATIONS
REPLICA TECHNIQUES