Towards native-state imaging in biological context in the electron microscope

Journal of Chemical Biology

Hannah E. J. Armer¹  Anne E. Weston¹  Lucy M. Collinson¹ 

[1] Electron Microscopy Unit, London Research Institute, Cancer Research UK, 44 Lincoln’s Inn Fields, London, WC2A 3PX UK

关键词: Transmission electron microscopy;    Scanning electron microscopy;    Artifacts;    Native state;    Cryopreparation;    Cryo-EM;    ESEM;    Correlative;    Volume EM;    FIB/SEM;    SBF/SEM;   

DOI  :  10.1007/s12154-009-0033-7

学科分类：分子生物学，细胞生物学和基因

来源: Springer

【 摘 要 】

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.

【 授权许可】

Unknown