Scaling down the bioimaging of metals by laser microdissection inductively coupled plasma mass spectrometry (LMD-ICP-MS)

Becker, J. Sabine; Niehren, Stefan; Matusch, Andreas; Wu, Bei; Hsieh, H. F.; Kumtabtim, Usarat; Hamester, M.; Plaschke-Schlütter, A.; Salber, Dagmar

doi:10.1016/j.ijms.2010.03.013

Cited by 30 publications

(27 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A high resolution can be achieved using a laser beam with a smaller spot size, although this may lead to insufficient energy focused on the sample and, hence, limited ablation of the material. A laser microdissection (LMD) instrument can substitute a laser ablation system to substantially enhance the spatial resolution [26].…”

Section: Trends In Plant Sciencementioning

confidence: 99%

“…NanoSIMS has been used to reveal the uptake and competition between rhizosphere microorganisms and plant roots for 15 N labeled ammonium [55,56]. Pulsing cut stems of bean (Phaseolus vulgaris) plants with 26 Mg, 41 K, 44 Ca, and H 2 18 O and using cryo-SIMS to image their distribution at different times revealed a rapid exchange of cations between xylem vessels and the adjacent xylem parenchyma cells, and a slow exchange with cambium and phloem; the three cations also exhibited different exchange kinetics [57,58]. In these studies, samples were shock-frozen in melting propane, fractured, and analyzed under cryo-conditions, thus preserving the cellular structure and in situ element distribution.…”

Section: Trends In Plant Sciencementioning

confidence: 99%

See 1 more Smart Citation

Imaging element distribution and speciation in plant cells

Zhao

Moore

Lombi

et al. 2014

Trends in Plant Science

138

103

View full text Add to dashboard Cite

To maintain cellular homeostasis, concentrations, chemical speciation, and localization of mineral nutrients and toxic trace elements need to be regulated. Imaging the cellular and subcellular localization of elements and measuring their in situ chemical speciation are challenging tasks that can be undertaken using synchrotronbased techniques, such as X-ray fluorescence and X-ray absorption spectrometry, and mass spectrometry-based techniques, such as secondary ion mass spectrometry and laser-ablation inductively coupled plasma mass spectrometry. We review the advantages and limitations of these techniques, and discuss examples of their applications, which have revealed highly heterogeneous distribution patterns of elements in different cell types, often varying in chemical speciation. Combining these techniques with molecular genetic approaches can unravel functions of genes involved in element homeostasis. Spatial and chemical information of mineral element homeostasisPlants take up a range of mineral elements from the soil, some of which are essential for growth, whereas others are non-essential [1]. Deficiencies of essential elements are a major limiting factor for crop production in many areas worldwide, whereas excessive accumulation of both essential and non-essential elements can lead to phytotoxicity [1]. Accumulation of some elements, such as cadmium (Cd) [2] and arsenic (As) [3], in the edible parts of crops may pose a significant risk to human health well before phytotoxicity occurs. By contrast, there is a need to increase essential micronutrients, such as iron (Fe) and zinc (Zn), in plantbased foods to alleviate their deficiencies in humans [4]. Plant nutrition research aims to understand how minerals are acquired, transported, distributed, stored, and used in plants. This knowledge is important not only for sustainable agricultural production but also for ensuring the nutritional quality and safety of agricultural products [5].Analyses of total elemental concentrations can now be performed using high-throughput platforms to reveal the ionomic profile (see Glossary) of plant tissues [6]. Although the total concentrations of minerals can provide information about the capacity for uptake and translocation, it is well recognized that minerals are distributed heterogeneously across different cell types [7]. Not only may the total concentrations vary at the tissue, cellular, and subcellular scales but also the chemical speciation of minerals may vary. This spatial information is crucial for understanding the homeostasis of minerals, particularly how different cell types and, fundamentally, different genes function in controlling the distribution, complexation, and storage of minerals, and how these processes vary among diverse plant species in the ecophysiological context.A range of techniques are available for mapping element distribution at various spatial scales. Traditional methods, such as energy-dispersive X-ray microanalysis (EDX) and proton (particle)-induced X-ray emission (PIXE), have been u...

show abstract

Section: Trends In Plant Sciencementioning

confidence: 99%

Section: Trends In Plant Sciencementioning

confidence: 99%

Imaging element distribution and speciation in plant cells

Zhao

Moore

Lombi

et al. 2014

Trends in Plant Science

138

103

View full text Add to dashboard Cite

show abstract

“…Compared with the previous study, 18 the ablation of the material in a 30-mmthick brain standard tissue was more efficient and a high ion intensity of elements was monitored in ICP-MS. For the first time, imaging of a biological tissue was achieved with spatial resolution of 30 mm, 15 mm and 8 mm, respectively, by the novel LMD-ICP-MS imaging technique. Moreover, images of Mg, K, Fe and P with a lateral resolution of 4 mm was achieved in a small region of a mouse brain section by imaging LMD-ICP-MS. Quantitative LMD-ICP-MS was developed by using the quantification procedures applied in routine LA-ICP-MS imaging via calibration curves from the matrix-matched brain standards.…”

Section: Resultsmentioning

confidence: 92%

Mass spectrometric imaging of elements in biological tissues by new BrainMet technique—laser microdissection inductively coupled plasma mass spectrometry (LMD-ICP-MS)

Niehren²,

Becker

2011

J. Anal. At. Spectrom.

View full text Add to dashboard Cite

Bioimaging of elements in biological tissues is a fast growing technology in the life sciences since it provides direct information on the metal distribution which is involved in metabolism in all kinds of organisms. In the present study, we proposed a novel mass spectrometric imaging technique, laser microdissection inductively coupled plasma mass spectrometry (LMD-ICP-MS), which is intended to investigate elemental distribution in small areas of biological tissue with high spatial resolution down to the low-micrometre range and below. The LMD-ICP-MS technique combines a laser microdissection apparatus (LMD) with sensitive quadrupole-based inductively coupled plasma mass spectrometry (ICP-QMS) to utilize the tightly focused laser beam in LMD for the ablation of sample materials which then are analyzed by ICP-QMS. The laser beam with a 1 mm spot diameter was used to ablate the material from a Cu-spiked brain tissue slice, and corresponding ion intensities of elements of interest (for instance, Cu + , 64 Zn + ) in the scanned regions was revealed by LMD-ICP-MS imaging with a spatial resolution of 30 mm, 15 mm, and 8 mm, respectively. Additionally, images of P, Mg, K and Fe with a lateral resolution of 4 mm in a region of a mouse brain slice were obtained by imaging LMD-ICP-MS. Quantification in LMD-ICP-MS was possible via calibration curves generated by a set of synthetic brain standards similar to the procedure in laser ablation ICP-MS. Further studies with respect to the performance of LMD-ICP-MS and the optimization of experimental parameters when using laser beams with smaller spot sizes in LMD will be carried out in developing the LMD-ICP-MS imaging technique.

show abstract

“…For the normalization of topographic differences or differences in tissue hardness, an element already present in the tissue can be used. Carbon ( 13 C) has been used frequently in other LA-ICP-MS studies (Becker et al, 2010a(Becker et al, , 2010b. In our root sections, the signal from 13 C was very weak, which made this approach unreliable.…”

Section: Sample Preparation and La-icp-ms Analysismentioning

confidence: 88%

“…New LA designs, however, like the near-field LA type, can go below 5 mm, and substitution of the LA unit with a laser microdissection unit has been used to improve resolution down to 1 mm (Becker et al, 2010a).…”

Section: Discussionmentioning

confidence: 99%

Multi-element bioimaging of Arabidopsis thaliana roots

et al. 2016

View full text Add to dashboard Cite

(J.K.S.); 0000-0003-2020-1902 (S.H.).Better understanding of root function is central for the development of plants with more efficient nutrient uptake and translocation. We here present a method for multielement bioimaging at the cellular level in roots of the genetic model system Arabidopsis (Arabidopsis thaliana). Using conventional protocols for microscopy, we observed that diffusible ions such as potassium and sodium were lost during sample dehydration. Thus, we developed a protocol that preserves ions in their native, cellular environment. Briefly, fresh roots are encapsulated in paraffin, cryo-sectioned, and freeze dried. Samples are finally analyzed by laser ablation-inductively coupled plasma-mass spectrometry, utilizing a specially designed internal standard procedure. The method can be further developed to maintain the native composition of proteins, enzymes, RNA, and DNA, making it attractive in combination with other omics techniques. To demonstrate the potential of the method, we analyzed a mutant of Arabidopsis unable to synthesize the metal chelator nicotianamine. The mutant accumulated substantially more zinc and manganese than the wild type in the tissues surrounding the vascular cylinder. For iron, the images looked completely different, with iron bound mainly in the epidermis of the wild-type plants but confined to the cortical cell walls of the mutant. The method offers the power of inductively coupled plasma-mass spectrometry to be fully employed, thereby providing a basis for detailed studies of ion transport in roots. Being applicable to Arabidopsis, the molecular and genetic approaches available in this system can now be fully exploited in order to gain a better mechanistic understanding of these processes.Investigations of the localization of inorganic elements in young plant roots may answer a range of important and unresolved questions with respect to root functionality and plant nutrient transport. To date, our understanding of how plants control the radial root transport of essential plant nutrients and toxic elements is mainly circumstantial, relying on changes in shoot or shoot-to-root concentration ratios or analyses of xylem sap composition. Roots of Arabidopsis (Arabidopsis thaliana) have a simple cellular organization and are unrivaled in their ability to be imaged by confocal microscopy, as they are very thin (diameter approximately 120 mm) and have a low background fluorescence. This has led to an amazingly detailed understanding of the growth and development of roots. Unfortunately, the fragile nature of these roots constitutes a major challenge when trying to understand the processes that drive nutrient uptake at the same level of detail. The method we present here for element bioimaging of Arabidopsis roots is a critical step in utilizing the potential of combining targeted genetic modifications and bioimaging at the cellular level in order to unravel the complexities of how roots selectively acquire and translocate mineral nutrients from the soil.The uptake and radial tr...

show abstract

Scaling down the bioimaging of metals by laser microdissection inductively coupled plasma mass spectrometry (LMD-ICP-MS)

Cited by 30 publications

References 25 publications

Imaging element distribution and speciation in plant cells

Imaging element distribution and speciation in plant cells

Mass spectrometric imaging of elements in biological tissues by new BrainMet technique—laser microdissection inductively coupled plasma mass spectrometry (LMD-ICP-MS)

Multi-element bioimaging of Arabidopsis thaliana roots

Contact Info

Product

Resources

About