Abstract
The techniques of electron energy-loss spectroscopy (EELS) and energy-filtered TEM (EFTEM) are routinely applied in the physical sciences to map the distribution of elements at the nanoscale. EELS can also provide details of the bonding/valence of elements through variations in the fine structure of elemental peaks in the spectrum. While applications of these techniques in biology are less prevalent, their ability to detect both the light elements (e.g., C, N, O, P, S) that form the building blocks of biological systems and heavier elements (e.g., metals) makes them potentially important techniques for investigating local chemical variations in tissues and cells. Successful application of EELS and EFTEM in biology requires both an understanding of the techniques themselves and expertise in specimen preparation. Care must be taken to avoid the diffusion of elements during the preparation process to avoid artifacts in the resulting element maps. The power of the techniques is demonstrated here using tissue from a marine mollusc (chiton).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Brydson R (2001) Electron energy loss spectroscopy. BIOS Scientific Publishers Limited, Oxford
Keast VJ, Scott AJ, Brydson R et al (2001) Electron energy-loss near-edge structure—a tool for the investigation of electronic structure on the nanometre scale. J Microsc (Oxford) 203:135–175
Thomas PJ, Midgley PA (2002) An introduction to energy-filtered transmission electron microscopy. Topics Catalysis 21:109–138
Verbeek J, Van Dyck D, Van Tendeloo G (2004) Energy-filtered transmission electron microscopy: an overview. Spectrochimica Acta Part B 59:1529–1534
Pennycook SJ (2012) Energy-filtered transmission electron microscopy: an overview. MRS Bull 37:943–951
Aranova MA, Kim YC, Zhang G et al (2007) Quantification and thickness correction of EFTEM phosphorus maps. Ultramicroscopy 107:232–244
Arsenault AL, Ottensmeyer FP (1983) Quantitative spatial distributions of calcium, phosphorus, and sulfur in calcifying epiphysis by high resolution electron spectroscopic imaging. Proc Natl Acad Sci U S A 80:1322–1326
Leapman RD, Jarnik M, Stevens AC (1997) Spatial distributions of sulfur-rich proteins in cornifying epithelia. J Struct Biol 120:168–179
Clode PL, Saunders M, Ludwig M et al (2009) Urate deposits in symbiotic marine algae. Plant Cell Environ 32:170–177
Lipovsek S, Letofsky-Papst I, Hofer F et al (2012) Application of analytical electron microscopic methods to investigate the function of spherites in the midgut of the larval antlion Euroleon nostras (Neuroptera: Myrmeleontidae). Microsc Res Tech 75:397–407
Aranova MA, Kim YC, Pivovarova NB et al (2009) Quantitative EFTEM mapping of near physiological calcium concentrations in biological specimens. Ultramicroscopy 109:201–212
Shaw JA, Clode PL, Brooker LR et al (2009) The chiton stylus canal: an element delivery pathway for tooth cusp biomineralization. J Morphol 270:588–600
Shaw JA, Clode PL, Brooker LR et al (2009) Ultrastructure of the epithelial cells associated with tooth biomineralization in the chiton Acanthopleura hirtosa. Microsc Microanal 15:154–165
Saunders M, Kong C, Shaw JA et al (2009) Characterization of biominerals in the radula teeth of the chiton, acanthopleura hirtosa. J Struct Biol 167:55–61
Usher KM, Shaw JA, Kaksonen AH et al (2010) Elemental composition of extracellular polymeric substances and granules in chalcopyrite bioleaching microbes. Hydrometallurgy 104:376–381
Saunders M, Kong C, Shaw JA et al (2011) Matrix-mediated biomineralization in marine molluscs: a combined TEM and FIB approach. Microsc Microanal 17:220–225
Treiber CD, Salzer M, Riegler J et al (2012) Clusters of iron rich cells in the upper beak of the pigeon are macrophages not magnetosensitive neurons. Nature 484:367–370
Chan EPH, Mhawi A, Clode PL et al (2009) Effects of titanium (IV) ions on human dendritic cells. Metallomics 1:166–174
Wedlock L, Kilburn M, Cliff JB et al (2011) Visualizing gold inside tumor cells following treatment with an antitumor gold(I) complex. Metallomics 3:917–925
Davis J, Heng YM, Barfels MMG et al (2000) Localization of chromophore absorption signals in TEM with an improved prism-mirror-prism filter. J Electron Microsc 49:629–639
Kothleitner G, Hofer F (1998) Optimization of the signal to noise ratio in EFTEM elemental maps with regard to different ionization edge types. Micron 29:349–357
Aranova MA, Leapman RD (2012) Development of electron energy-loss spectroscopy in the biological sciences. MRS Bulletin 37:53–62
Echlin P (1992) Low-temperature microscopy and analysis. Plenum Press, London
Ingram P, Shelburne JD, Roggli VL et al (1999) Biomedical applications of microprobe analysis. Academic, London
Steinbrecht RA, Müller M (1987) Freeze-substitution and freeze-drying. In: Steinbrecht RA, Zierold K (eds) Cryotechniques in biological electron microscope. Springer, Berlin, pp 149–172
Egerton RF (1996) Electron energy-loss spectroscopy in the electron microscope, 2nd edn. Plenum Press, New York
Muller-Reichert T (ed) (2010) Electron microscopy of model systems. Academic, San Diego, USA
McDonald KL, Webb RI (2011) Freeze substitution in 3 hours or less. J Microsc (Oxford) 243:227–233
Grogger W, Schaffer B, Krishnan KM et al (2003) Energy-filtering TEM at high magnification: spatial resolution and detection limits. Ultramicroscopy 96:481–489
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media, New York
About this protocol
Cite this protocol
Saunders, M., Shaw, J.A. (2014). Biological Applications of Energy-Filtered TEM. In: Kuo, J. (eds) Electron Microscopy. Methods in Molecular Biology, vol 1117. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-776-1_31
Download citation
DOI: https://doi.org/10.1007/978-1-62703-776-1_31
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-775-4
Online ISBN: 978-1-62703-776-1
eBook Packages: Springer Protocols