Two-Dimensional Mapping of Copper and Zinc in Liver Sections by Laser Ablation–Inductively Coupled Plasma Mass Spectrometry

Kindness, Andrew; Sekaran, Chandra N.; Feldmann, Jörg

doi:10.1373/clinchem.2003.022046

Cited by 133 publications

(110 citation statements)

References 20 publications

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“…A more sensitive examination of the total zinc content was subsequently possible via atomic emission, absorption or fluorescence spectroscopy, but also with other techniques such as inductively coupled plasma mass spectrometry [7,11,34] and laser ablation-inductively coupled plasma mass spectrometry. The latter even allow a two-dimensional mapping of the distribution of zinc in organic samples [35].…”

Section: Measurement Of Total Zincmentioning

confidence: 99%

Parameters Influencing Zinc in Experimental Systems in Vivo and in Vitro

Ollig

Kloubert

Weßels

et al. 2016

Metals

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Abstract:In recent years, the role of zinc in biological systems has been a subject of intense research. Despite wide increase in our knowledge and understanding of zinc homeostasis, numerous questions remain to be answered, encouraging further research. In particular, the quantification of intracellular zinc ions and fluctuation, as well as the function of zinc in signaling processes are being intensely investigated. The determination of free intracellular zinc ions is difficult and error-prone, as concentrations are extremely low (in the pico-to nanomolar range), but techniques exist involving fluorescent probes and sensors. In spite of zinc deficiency being accepted as a global problem, causing death and disease worldwide, to date there are no markers to reliably assess a person's zinc status. This review summarizes the difficulties and major pitfalls when working with zinc in in vitro and in vivo research. Additionally, it specifies important aspects for zinc substitution and supplementation, including the bioavailability of zinc and its intestinal absorption. In particular, it is intended to help researchers with yet minor experience working with zinc efficiently set up experiments and avoid commonly occurring mistakes, starting with the choice and preparation of reagents and instrumentation, and concluding with possibilities for measuring the status of zinc in humans.

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Section: Measurement Of Total Zincmentioning

confidence: 99%

Parameters Influencing Zinc in Experimental Systems in Vivo and in Vitro

Ollig

Kloubert

Weßels

et al. 2016

Metals

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“…Another option is the use of a cryo-cell for ablation of a frozen gel as we described for the ablation of biological tissue [35,36]. The main problem encountered was that of irregular ice crystal formation on the surface and within the gel, whether the gel was pre-frozen in liquid nitrogen or frozen in the ablation chamber, which hindered ablation.…”

Section: Generalmentioning

confidence: 99%

Evaluation of gel electrophoresis conditions for the separation of metal‐tagged proteins with subsequent laser ablation ICP‐MS detection

et al. 2009

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Although laser ablation (LA)-ICP-MS has been reported for the determination of metalloproteins separated by gel electrophoretic techniques (GE), systematic studies that define the conditions essential for successful measurements are still scarce. In this paper we present the results of our studies of basic conditions for the effective application of GE-LA-ICP-MS for the separation of metal-binding proteins, focusing on their stability during GE and post-separation gel treatment. The stability of metal-protein complexes (haemoglobin, myoglobin, superoxide dismutase, carbonic anhydrase, transferrin, albumin, cytochrome c) during GE is dependent on the nature of the metal-protein interaction and the principle of separation. We have observed that non-denaturing GE is a suitable separation technique for most metal-protein complexes (e.g. Zn in carbonic anhydrase and Fe in Tf and myoglobin were quantitatively recovered in a spiked liver cytosol), whereas separation by denaturing GE strongly impaired the stability of the complexes. Equally important is the post-separation treatment of the gel to enable successful detection of the metal. LA-ICP-MS requires drying of the gel without loss of protein-bound metal or cracking of the gel. This was successfully achieved using glycerol followed by heating. We demonstrate that staining of the gel prior to LA-ICP-MS using silver or Coomassie blue is not recommended, since most protein-bound metal is lost during the staining procedure. Furthermore it has been shown that only line scanning with a speed of less than 30 microm/s can reliably distinguish between lines 1 mm apart, while raster spot analysis carries the risk of misinterpretation due to contamination in/on inhomogeneous gels.

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“…However, they showed low precision or poor detection limits. Using laser ablation-inductively coupled plasma mass spectrometry an elemental mapping of Cu and Zn was achieved in sheep liver slices [17].…”

Section: Introductionmentioning

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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Two-Dimensional Mapping of Copper and Zinc in Liver Sections by Laser Ablation–Inductively Coupled Plasma Mass Spectrometry

Cited by 133 publications

References 20 publications

Parameters Influencing Zinc in Experimental Systems in Vivo and in Vitro

Parameters Influencing Zinc in Experimental Systems in Vivo and in Vitro

Evaluation of gel electrophoresis conditions for the separation of metal‐tagged proteins with subsequent laser ablation ICP‐MS detection

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

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