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Developing post embedding staining technique for EDX analysis

Lusseveld, Jarnick (2018) Developing post embedding staining technique for EDX analysis. Master's Research Project 2, Molecular Biology and Biotechnology (2016-2019).

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Thesis Cell biology Final (J.H.Lusseveld).pdf
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Electron microscopy is a powerful tool which allows great insight in biological processes because of its high resolution. Images in scanning electron microscopy (SEM) are created by scanning a sample with an electron beam and counting the number of backscattered electrons for each pixel (Gunther Rudenberg & Rudenberg, 2010). Contrast arises by heavier elements causing greater scattering of electrons. It is however still difficult in many cases to identify cellular structures, because the obtained image consists of grey values. Additional information can be added to SEM images by collecting x-rays emitted by the sample when the electron beam moves over the sample(Scotuzzi et al., 2017). Identifying specific x-rays during SEM is called energy dispersive x-ray spectroscopy (EDX)(Falke et al., n.d.). Elements present in the sample will emit characteristic x-rays, which can then be used to form element maps. Indigenous elements can then be detected and cellular features easier identified with more certainty. Moreover, it is possible to add endogenous elements to the sample that have a specific affinity for cellular components or cell types and using EDX to analyze the sample. The objective of this thesis is to identify metallic stains that provide additional information about cellular features using EDX. To label cellular features with different metals, rat pancreas tissue was stained with different metals under multiple pH conditions and analyzed using a SEM microscope with an x-ray detector. Several metals have been identified to stain the nucleus (heterochromatin) and granules in endocrine and exocrine cells of the rat pancreas. This is the case for samarium acetate, cobalt chloride, iridium bromide, and silver lactate. Additional experiments are still necessary to confirm and optimize the described metal staining techniques in this project.

Item Type: Thesis (Master's Research Project 2)
Supervisor name: Giepmans, B.N.G.
Degree programme: Molecular Biology and Biotechnology (2016-2019)
Thesis type: Master's Research Project 2
Language: English
Date Deposited: 06 Sep 2018
Last Modified: 06 Sep 2018 10:38

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