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Electron tomography of HEK293T cells using scanning electron microscope-based scanning transmission electron microscopy

, : Electron tomography of HEK293T cells using scanning electron microscope-based scanning transmission electron microscopy. Microscopy and Microanalysis 18(5): 1037-1042

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.


PMID: 23026379

DOI: 10.1017/S1431927612001158

Other references

Pascual, A.C.; Gaveglio, V.L.; Giusto, N.M.; Pasquaré, S.J., 2013: Aging modifies the enzymatic activities involved in 2-arachidonoylglycerol metabolism. One of the principal monoacylglycerol (MAG) species in animal tissues is 2-arachidonoylglycerol (2-AG), and the diacylglycerol lipase (DAGL) pathway is the most important 2-AG biosynthetic pathway proposed to date. Lysophosphatidate phosphatase (L...

Lomberg, H.; Cedergren, B.; Leffler, H.; Nilsson, B.; Carlström, A.S.; Svanborg-Edén, C., 1986: Influence of blood group on the availability of receptors for attachment of uropathogenic Escherichia coli. Escherichia coli strains with defined receptor specificity were used as probes to analyze the individual variation in host cell receptors with respect to blood groups. The adhesins were initially characterized as mannose sensitive (MS), mannose re...

Amano, Y.; Yamashita, H.; Koshino, K.; Ohshima, T.; Miwa, H.; Iwakiri, R.; Fujimoto, K.; Manabe, N.; Haruma, K.; Kinoshita, Y., 2008: Does magnifying endoscopy improve the diagnosis of erosive esophagitis?. Background and Aims: Low-grade erosive esophagitis (i.e. Los Angeles grade A) is the most predominant type of esophagitis in Japan. It is unclear whether all the mucosal breaks detected by conventional endoscopy are indicative of esophageal mucos...

Cha, S.; Johnson, G.; Wadghiri, Y.Zaim.; Jin, O.; Babb, J.; Zagzag, D.; Turnbull, D.H., 2003: Dynamic, contrast-enhanced perfusion MRI in mouse gliomas: correlation with histopathology. The aim of this study was to develop an MRI protocol to evaluate the growth and vascularity of implanted GL261 mouse gliomas on a 7T microimaging system. Both conventional T(1)- and T(2)-weighted imaging and dynamic, contrast-enhanced T(2)*-weight...

Katalymov, L.L., 1973: Method of determining trace changes in excitability. Nauchnye Doklady Vysshei Shkoly. Biologicheskie Nauki 120(12): 116-124

Gray, J.A., 1997: Underpricing and overexploitation of tropical forests: forest pricing in the management, conservation, and preservation of tropical forests. Forest pricing policies for tropical timber and forest concessions can play an important role in the management, conservation, and preservation of tropical forests. The paper draws on regional studies by the author and others of forest revenue sys...

Nakata, Y., 1974: The role of hypoxia and pH changes in the cardiac arrhythmias. Kokyu to Junkan. Respiration & Circulation 22(4): 327-332

Heyburn Ross G.; Bowers D., 2004: Depth determination of earthquakes near the Chinese test site using the F trace and associated probability. Seismological Research Letters 75(2): 252

Skosey J.L., 1985: Immunobiological functions of articular chondrocytes. Arthritis & Rheumatismsuppl: S31

Kushinov N.V., 1985: The Teplovian Suite of the Belozerian Synclinore. Geologicheskiy Zhurnal (Kiev, 1968) 45(4): 71-74