Dual-axis electron tomography of biological specimens: Extending the limits of specimen thickness with bright-field STEM imaging.
J Struct Biol
; 174(1): 107-14, 2011 Apr.
Article
em En
| MEDLINE
| ID: mdl-21055473
ABSTRACT
The absence of imaging lenses after the specimen in the scanning transmission electron microscope (STEM) enables electron tomography to be performed in the STEM mode on micrometer-thick plastic-embedded specimens without the deleterious effect of chromatic aberration, which limits spatial resolution and signal-to-noise ratio in conventional TEM. Using Monte Carlo calculations to simulate electron scattering from gold nanoparticles situated at the top and bottom surfaces of a plastic section, we assess the optimal acquisition strategy for axial bright-field STEM electron tomography at a beam-energy of 300keV. Dual tilt-axis STEM tomography with optimized axial bight-field detector geometry is demonstrated by application to micrometer-thick sections of beta cells from mouse pancreatic islet. The quality of the resulting three-dimensional reconstructions is comparable to that obtained from much thinner (0.3-micrometer) sections using conventional TEM tomography. The increased range of specimen thickness accessible to axial STEM tomography without the need for serial sectioning enables the 3-D visualization of more complex and larger subcellular structures.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Microscopia Eletrônica de Transmissão e Varredura
/
Tomografia com Microscopia Eletrônica
Tipo de estudo:
Health_economic_evaluation
Limite:
Animals
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article