Principles and instrumentation

Ingram, Peter; Shelburne, John D.; LeFurgey, Ann

doi:10.1016/b978-012371020-8/50003-2

Cited by 28 publications

(8 citation statements)

References 152 publications

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“…925; Gatan, Inc., Pleasanton, CA), a scanning device, additional hard X-ray aperture, a collimated 30-mm 2 Si(Li) energy-dispersive X-ray detector (Oxford Instruments America, Palo Alto, CA), and a pulse processor; the scanning and multichannel analyses were conducted with an X-ray pulse processor (4pi Analysis spectral engine; 4pi Analysis, Inc., Durham, NC). Operating parameters and strategies for obtaining quantitative X-ray images were implemented as described extensively elsewhere (19,(25)(26)(27). Basically, 128-by 128-pixel spectral data sets were obtained at approximately ϫ10,000 magnification (1 pixel ϭ 0.2 m 2 ) with a beam current of approximately 1 nA and a dwell time of 4 s per pixel, with automation (19) and quantitation (27) carried out as previously described.…”

Section: Methodsmentioning

confidence: 99%

“…Operating parameters and strategies for obtaining quantitative X-ray images were implemented as described extensively elsewhere (19,(25)(26)(27). Basically, 128-by 128-pixel spectral data sets were obtained at approximately ϫ10,000 magnification (1 pixel ϭ 0.2 m 2 ) with a beam current of approximately 1 nA and a dwell time of 4 s per pixel, with automation (19) and quantitation (27) carried out as previously described. The elemental contents of whole cells and cell regions (i.e., vacuoles or nonvacuolar cytoplasm) were determined by drawing around, i.e., masking, each cell or individual vacuole present; with this method, the cytoplasmic compartment included all structures except the vacuole, i.e., the endoplasmic reticulum (ER), Golgi body, nucleus, mitochondria, and lysosomes.…”

Section: Methodsmentioning

confidence: 99%

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Saccharomyces cerevisiae Vacuole in Zinc Storage and Intracellular Zinc Distribution

et al. 2007

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Previous studies of the yeast Saccharomyces cerevisiae indicated that the vacuole is a major site of zinc storage in the cell. However, these studies did not address the absolute level of zinc that was stored in the vacuole nor did they examine the abundances of stored zinc in other compartments of the cell. In this report, we describe an analysis of the cellular distribution of zinc by use of both an organellar fractionation method and an electron probe X-ray microanalysis. With these methods, we determined that zinc levels in the vacuole vary with zinc status and can rise to almost 100 mM zinc (i.e., 7 ؋ 10 8 atoms of vacuolar zinc per cell). Moreover, this zinc can be mobilized effectively to supply the needs of as many as eight generations of progeny cells under zinc starvation conditions. While the Zrc1 and Cot1 zinc transporters are essential for zinc uptake into the vacuole under steady-state growth conditions, additional transporters help mediate zinc uptake into the vacuole during "zinc shock," when zinc-limited cells are resupplied with zinc. In addition, we found that other compartments of the cell do not provide significant stores of zinc. In particular, zinc accumulation in mitochondria is low and is homeostatically regulated independently of vacuolar zinc storage. Finally, we observed a strong correlation between zinc status and the levels of magnesium and phosphorus accumulated in cells. Our results implicate zinc as a major determinant of the ability of the cell to store these other important nutrients.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Saccharomyces cerevisiae Vacuole in Zinc Storage and Intracellular Zinc Distribution

et al. 2007

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“…The microscope was fitted with custom-made LN 2 -cooled anticontaminators and a LaB 6 filament. Analyses were carried out by elemental imaging using the multispectral analysis program ImagNspect (Ingram et al, 1999), at 120·kV with a beam diameter of <90·nm and a beam current of 5ϫ10 -10 ·A. Peak integral and quantitative images were obtained with a resolution of 128ϫ128·pixels and a dwell time of 3·s·pixel -1 .…”

Section: X-ray Microanalysismentioning

confidence: 99%

Electron and ion microprobe analysis of calcium distribution and transport in coral tissues

Marshall

Clode

Russell

et al. 2007

Journal of Experimental Biology

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“…The specimens were analyzed at room temperature in a carbon holder tilted at 25° towards the detector. Elemental imaging was carried out using the multispectral analysis program ImagNspect (Ingram et al 1999). Elemental maps were obtained with a resolution of 128 £ 128 pixels and a dwell time of 3 s per pixel.…”

Section: Energy-dispersive X-ray Microanalysismentioning

confidence: 99%

Cellular and subcellular compartmentation of Ni in the Eurasian serpentine plants Alyssum bracteatum, Alyssum murale (Brassicaceae) and Cleome heratensis (Capparaceae)

Asemaneh

Ghaderian

Crawford

et al. 2006

Planta

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This study investigated the cellular and subcellular compartmentation of Ni in the Eurasian serpentine species Alyssum murale, Alyssum bracteatum and Cleome heratensis and a non-serpentine population of A. murale (as a control) grown in hydroponic culture. Plant growth responses and Ni uptake clearly revealed the higher Ni tolerance of serpentine plants than the non-serpentine plants. Serpentine A. murale and A. bracteatum grew better at elevated (0.01 mM) Ni in the nutrient solution, supporting the view that the Ni hyperaccumulators have a higher requirement for Ni than normal plants. Low shoot Ni content of C. heratensis in response to the high Ni treatments indicated that this species employs an avoidance strategy for Ni tolerance. Energy-dispersive X-ray microanalysis showed that Ni was highly concentrated in the cell walls and cell lumen, most likely the vacuoles, of leaf epidermis of A. murale and A. bracteatum rather than in the mesophyll cells. EDX spectra from leaves of the non-serpentine A. murale suggested that Ni accumulated in both epidermal and mesophyll cells but not in the epidermal cell walls. Growth reduction and Ni toxicity in plants of the non-serpentine A. murale could be due to accumulation of Ni in the lumen of leaf mesophyll cells. Our data suggest that cellular and subcellular compartmentation are both possible mechanisms for Ni tolerance employed by the serpentine A. murale and A. bracteatum.

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Principles and instrumentation

Cited by 28 publications

References 152 publications

Saccharomyces cerevisiae Vacuole in Zinc Storage and Intracellular Zinc Distribution

Saccharomyces cerevisiae Vacuole in Zinc Storage and Intracellular Zinc Distribution

Electron and ion microprobe analysis of calcium distribution and transport in coral tissues

Cellular and subcellular compartmentation of Ni in the Eurasian serpentine plants Alyssum bracteatum, Alyssum murale (Brassicaceae) and Cleome heratensis (Capparaceae)

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