Preparation of tissue samples for X-ray fluorescence microscopy

Chwiej, Joanna; Szczerbowska-Boruchowska, Magdalena; Lankosz, M.; Wójcik, Sławomir; Falkenberg, Gerald; Stęgowski, Z.; Setkowicz, Zuzanna

doi:10.1016/j.sab.2005.10.002

Cited by 69 publications

(63 citation statements)

References 23 publications

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“…Conversely, calcium and the major biologically-relevant transition metals (iron, copper and zinc) all showed increased concentrations within cells. Similar observations were made by Chwiej et al, 19 who used X-ray fluorescence spectroscopy to show differential influx and efflux of biologically-relevant elements in various areas of rat brain sections following formalin or paraformaldehyde fixation, with paraffin-embedded counterparts showing a marked decrease in nearly all measured elements. External contamination is clearly a point of consideration; when high-purity reagents are used, aldehyde fixation and sucrose cryoprotection resulted in loss of most biologically-relevant metals.…”

Section: Developing An Ihc-chemical Imaging Experimentssupporting

confidence: 82%

A guide to integrating immunohistochemistry and chemical imaging

et al. 2018

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Chemical imaging provides new insight into the fundamental atomic, molecular, and biochemical composition of tissue and how they are interrelated in normal physiology. Visualising and quantifying products of pathogenic reactions long before structural changes become apparent also adds a new dimension to understanding disease pathogenesis. While chemical imaging in isolation is somewhat limited by the nature of information it can provide (e.g. peptides, metals, lipids, or functional groups), integrating immunohistochemistry allows simultaneous, targeted imaging of biomolecules while also mapping tissue composition. Together, this approach can provide invaluable information on the inner workings of the cell and the molecular basis of diseases. Key learning points(1) Chemical imaging must be redefined to encompass all analytical techniques that can spatially map the chemical composition of a sample.

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Section: Developing An Ihc-chemical Imaging Experimentssupporting

confidence: 82%

A guide to integrating immunohistochemistry and chemical imaging

et al. 2018

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“…However, for comparison of absolute concentration values with other studies it should be noticed that the samples used in our study were chemically fixated and embedded in paraffin. It is known that this preparation can change the concentration of light elements significantly [42]. Potassium, for example, is almost completely eliminated.…”

Section: Haemachromatosismentioning

confidence: 99%

Iron, copper, zinc and bromine mapping in cirrhotic liver slices from patients with hemochromatosis studied by microscopic synchrotron radiation X-ray fluorescence analysis in continuous scanning mode

Osterode¹,

Falkenberg²,

Höftberger³

et al. 2007

Spectrochimica Acta Part B: Atomic Spectroscopy

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Iron (Fe) and copper (Cu) are essential metals in physiological cell metabolism. While Fe is easy to determine biochemically in histological slices, Cu and zinc (Zn) distribution is frequently critical in confirming the presence of an overload in disturbed Fe/Cu metabolism. To analyze Fe, Cu and Zn in a near histological resolution, energy dispersive microscopic synchrotron radiation X-ray fluorescence was applied.In normal liver tissue, after fixation and imbedding in paraffin, mean Fe, Cu and Zn concentrations were 152 ± 54, 20.1 ± 4.3 and 88.919.5 μg/g sample weight, respectively. No substantial, characteristic differences in their distribution were found in the two-dimensional scans. In slices from patients with hemochromatosis mean Fe, Cu and Zn concentrations were 1102 ± 539, 35.9 ± 14.6 and 27.2 ± 6.7 μg/g sample weight, respectively. Additionally, a significant decrease in phosphorus and sulphur concentrations existed. An increased Cu around cirrhotic regenerations nodules is mostly associated with a lymphocytic infiltration in this region. Analyzing concentrations of Fe in different regions of the samples show a clear negative dependency between Fe and Cu, Cu and Zn, but a positive one between Fe and Zn.Conclusion: With a focal beam size of 15 μm in diameter a resolution of the elemental distribution was achieved which is widely comparable with stained histological slices (20× light microscope). The analysis of simultaneous determined elements reveals metabolic differences between Fe, Cu and Zn in liver tissue from patients with hemochromatosis.

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“…To our knowledge, it has never been used to investigate Cd at the L III -edge in plant samples. Artifacts resulting from the preparation and irradiation of biological samples have been observed [23], and in this study, we compared freeze-dried plant samples analyzed at ambient temperature and frozen hydrated samples analyzed at -170°C using a cryostat.…”

Section: Introductionmentioning

confidence: 99%