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Sample preparation for energy dispersive X-ray imaging of biological tissues

Authors :
Pirozzi, Nicole M
Kuipers, Jeroen
Giepmans, Ben N G
MullerReichert, T
Verkade, P
​Basic and Translational Research and Imaging Methodology Development in Groningen (BRIDGE)
Center for Liver, Digestive and Metabolic Diseases (CLDM)
Source :
Methods in Cell Biology ISBN: 9780128220580, CORRELATIVE LIGHT AND ELECTRON MICROSCOPY IV, 162, 89-114
Publication Year :
2021
Publisher :
Elsevier, 2021.

Abstract

Traditional electron microscopy (EM) can be complemented with analytical EM to increase objective sample information enabling feature identification. Energy dispersive X-ray (EDX) imaging provides semi-quantitative elemental composition of the sample with high spatial resolution (similar to 10nm) in ultrathin sections. However, EDX imaging of biological samples is still challenging as a routine method because many elements are at the detection limit for this technique. Moreover, samples undergo extensive preparation before analysis, which can introduce disruptive X-ray cross-talk or artifacts. EDX data can, for instance, be skewed by (i) osmium interference with endogenous phosphorus, (ii) chlorine present in EPONembedded tissues, (iii) lead interference with endogenous sulfur, and (iv) potential spectral overlaps with grid material, contrast agents, and the in-microscope sample holder. Here, we highlight how to circumvent these potential pitfalls and outline how we approach sample preparation and analysis for detection of different elements of interest. Utilization of wellconsidered a priori sample preparation techniques will best ensure conclusive EDX experiments.

Details

ISBN :
978-0-12-822058-0
ISBNs :
9780128220580
Database :
OpenAIRE
Journal :
Methods in Cell Biology ISBN: 9780128220580, CORRELATIVE LIGHT AND ELECTRON MICROSCOPY IV, 162, 89-114
Accession number :
edsair.doi.dedup.....ddc83e7f0417a96e404d0976606f183d