Towards native-state imaging in biological context in the electron microscope |
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Authors: | Anne E Weston Hannah E J Armer Lucy M Collinson |
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Institution: | (1) Electron Microscopy Unit, London Research Institute, Cancer Research UK, 44 Lincoln’s Inn Fields, London, WC2A 3PX, UK; |
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Abstract: | Modern cell biology is reliant on light and fluorescence microscopy for analysis of cells, tissues and protein localisation.
However, these powerful techniques are ultimately limited in resolution by the wavelength of light. Electron microscopes offer
much greater resolution due to the shorter effective wavelength of electrons, allowing direct imaging of sub-cellular architecture.
The harsh environment of the electron microscope chamber and the properties of the electron beam have led to complex chemical
and mechanical preparation techniques, which distance biological samples from their native state and complicate data interpretation.
Here we describe recent advances in sample preparation and instrumentation, which push the boundaries of high-resolution imaging.
Cryopreparation, cryoelectron microscopy and environmental scanning electron microscopy strive to image samples in near native
state. Advances in correlative microscopy and markers enable high-resolution localisation of proteins. Innovation in microscope
design has pushed the boundaries of resolution to atomic scale, whilst automatic acquisition of high-resolution electron microscopy
data through large volumes is finally able to place ultrastructure in biological context. |
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Keywords: | Transmission electron microscopy Scanning electron microscopy Artifacts Native state Cryopreparation Cryo-EM ESEM Correlative Volume EM FIB/SEM SBF/SEM |
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