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Electron Tomography of HEK293T Cells Using Scanning Electron Microscope-Based Scanning Transmission Electron Microscopy.
MedLine Citation:
PMID:  23026379     Owner:  NLM     Status:  Publisher    
Based on a scanning electron microscope operated at 30 kV with a homemade specimen holder and a multiangle solid-state detector behind the sample, low-kV scanning transmission electron microscopy (STEM) is presented with subsequent electron tomography for three-dimensional (3D) volume structure. Because of the low acceleration voltage, the stronger electron-atom scattering leads to a stronger contrast in the resulting image than standard TEM, especially for light elements. Furthermore, the low-kV STEM yields less radiation damage to the specimen, hence the structure can be preserved. In this work, two-dimensional STEM images of a 1-μm-thick cell section with projection angles between ±50° were collected, and the 3D volume structure was reconstructed using the simultaneous iterative reconstructive technique algorithm with the TomoJ plugin for ImageJ, which are both public domain software. Furthermore, the cross-sectional structure was obtained with the Volume Viewer plugin in ImageJ. Although the tilting angle is constrained and limits the resulting structural resolution, slicing the reconstructed volume generated the depth profile of the thick specimen with sufficient resolution to examine cellular uptake of Au nanoparticles, and the final position of these nanoparticles inside the cell was imaged.
Yun-Wen You; Hsun-Yun Chang; Hua-Yang Liao; Wei-Lun Kao; Guo-Ji Yen; Chi-Jen Chang; Meng-Hung Tsai; Jing-Jong Shyue
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-10-2
Journal Detail:
Title:  Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada     Volume:  -     ISSN:  1435-8115     ISO Abbreviation:  Microsc. Microanal.     Publication Date:  2012 Oct 
Date Detail:
Created Date:  2012-10-2     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9712707     Medline TA:  Microsc Microanal     Country:  -    
Other Details:
Languages:  ENG     Pagination:  1-6     Citation Subset:  -    
Research Center for Applied Sciences, Academia Sinica, Taipei 115, Taiwan.
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