To calculate PM2.5 mass concentrations in the exposure chambers, samples were collected weekly on Teflon filters (Teflon, 37 mm, 2 ɥm pore; Pall Life Sciences, Ann Arbor, MI, USA) and weighed before and after sampling in a temperature- and humidity-controlled weighing room using a microbalance (Mettle Toledo, Columbus, OH, USA). Samples collected from Teflon filters were wetted with ethanol and extracted in 1% nitric acid solution. The extraction solution was sonicated for 48 h in an ultrasonic bath and then allowed to passively acid digest for a minimum of 2 weeks. Sample extracts were then analyzed for a suite of trace elements using inductively coupled plasma-mass spectrometry (ELEMENT2, Thermo Finnigan, San Jose, CA, USA) as described [29 ].
Element 2
Manufactured by Thermo Fisher Scientific
Sourced in Germany, United States, Italy, France
The Element 2 is a high-performance, single-collector inductively coupled plasma mass spectrometer (ICP-MS) designed for elemental analysis. It provides accurate, sensitive, and reliable measurements across a wide range of sample types and concentrations.
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Lab products found in correlation
Milli q system, by Merck Group (3 mentions)
Nicolet is50, by Thermo Fisher Scientific (3 mentions)
Boron absorption standard solutions, by Merck Group (2 mentions)
Multiwave 3000, by Anton Paar (2 mentions)
Milli q reference a, by Merck Group (2 mentions)
Lsx 213 laser ablation system, by Thermo Fisher Scientific (2 mentions)
Teflon filter, by Pall Corporation (1 mentions)
Hpr 1000 6 m high pressure reactor, by Milestone (1 mentions)
Centrifuge 5424, by Eppendorf (1 mentions)
Normatom, by Avantor (1 mentions)
Tissueruptor, by Qiagen (1 mentions)
A3782 500mg, by Merck Group (1 mentions)
Fp20 0.45 ca s syringe filters, by Cytiva (1 mentions)
Human serum albumin (hsa), by Merck Group (1 mentions)
Ssm 5000a, by Shimadzu (1 mentions)
Total organic carbon analyser, by Shimadzu (1 mentions)
Toc vcph, by Shimadzu (1 mentions)
Ls 13 320, by Beckman Coulter (1 mentions)
Certipur, by Merck Group (1 mentions)
Dnase 1 solution, by Thermo Fisher Scientific (1 mentions)
106 protocols using element 2
1
PM2.5 Characterization in Exposure Chambers
2
Quantifying Lead Bioaccumulation in Zebrafish
3
Correlating Wood Localities Using U-Pb Zircon Geochronology
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Two tuff samples with abundant pumaceous fragments were collected to correlate two wood localities of the Colpamayo and San Miguel regions using U-Pb geochronology (Fig. 2 ). The sample from the Colpamayo region (STRI-MUSM 44449) was collected from a 60-cm conglomerate rich in pumaceous fragments, and the sample from the San Miguel region (STRI-MUSM 44448) was collected from a 50-cm-thick ashfall tuff. Zircon separates were performed by standard magnetic and gravity-based methods at the University of Rochester using a Frantz LB-1 separator and methylene iodide. Before U-Pb analysis by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), zircons were mounted in 1′ epoxy plugs and characterized by cathodoluminescence (CL) imaging using a JEOL JSM-7100 electron microscope equipped with a field-emission gun and a Deben Centaurus CL detector at the Mackay Microbeam Laboratory, University of Nevada, Reno. All U-Pb isotopic measurements were performed by LA-ICP-MS at the Arizona Laserchron Center, University of Arizona using a Photon Machines Analyte-G2 Excimer laser system coupled to a Thermo Finnigan Element2 single collector ICP-MS. Analytical methods for the U-Pb analyses are outlined in Ibáñez-Mejia et al. (42 ) and Pullen et al. (43 ) (see the Supplementary Materials for detailed methods and reference material).
4
Groundwater Arsenic Analysis Protocol
5
Rubidium Uptake in Doxorubicin-Treated Cells
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Seven thousand and fifty cells were plated in 10 cm Ø dishes. After 24 h, they are incubated with different concentrations of doxorubicin (from 0 to 5 µM) for a further 24 h. They were then incubated for 1 h with 0.12 mM RbCl in Earle’s Balanced Salt Solution (EBSS) at 37°C and 5% CO2. Following four washing steps with cold PBS to remove the extracellular RbCl, the cells were harvested. A Bradford assay was used to determine the protein concentration before acidic digestion. 200 μl of the cell suspension and 200 μl of HNO3 (70%) were subjected to acidic mineralization to oxidize the organic matter, solubilize all metals and simplify the matrix. The microwave (ETHOS UP Milestone, Bergamo, Italy) heating program consisted of a ramp to 150°C in 6 minutes, followed by 8 min to 150°C. After mineralization, sample volumes were brought to 3 ml with doubly deionized water. The quantification of Rb was performed via ICP-MS (Element-2; Thermo-Finnigan, Rodano (MI), Italy) analysis. The calibration curve was obtained using absorption standard solutions (Sigma-Aldrich) in the range 0.2–0.005 μg/mL.
6
Metal Fractionation in Synoviocytes
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Synoviocytes (5*105) were cultured with cytokines and metals being optionally added. Supernatants (2ml) and cells were uptake at the end-point (day 14 for metal cocktail and day 5 for Cd only) to analyze metal fractionation constants (KD). KD between cells and medium was calculated as KD = (Me/70Zn) cells/(Me/70Zn) medium. Values of KD of ~1 indicate an isotopic equilibrium, whereas KD>1 indicates that exchange reactions are still ongoing.
To analyze Cd kinetics and cell content, 2ml of supernatant were collected at 6, 12, 24, 48 hours. Cells were than washed with PBS and fresh complete DMEM was substituted for the Cd-enriched medium, to study the possible exit of Cd from cells. Two ml of medium were collected immediately after the wash, at 60, 72, 96 and 120 hours, while cells were collected and counted at the endpoint (120 hours). All the samples were then mineralized with HNO3 0.5N plus H2O2 (15–20%) at 100°C. Prior the analysis, the mineralized samples were re-dissolved in a 5% HNO3 solution in deionized water in an ultrasonic bath. Metal ions in the samples were then measured on a single collector ICP-MS platform ELEMENT 2 (Thermo Finnigan, Ringoes, NJ, USA) which allows metals to be measured at concentrations levels as low as 10−12 units, i.e. in the part per trillion (ppt) range.
To analyze Cd kinetics and cell content, 2ml of supernatant were collected at 6, 12, 24, 48 hours. Cells were than washed with PBS and fresh complete DMEM was substituted for the Cd-enriched medium, to study the possible exit of Cd from cells. Two ml of medium were collected immediately after the wash, at 60, 72, 96 and 120 hours, while cells were collected and counted at the endpoint (120 hours). All the samples were then mineralized with HNO3 0.5N plus H2O2 (15–20%) at 100°C. Prior the analysis, the mineralized samples were re-dissolved in a 5% HNO3 solution in deionized water in an ultrasonic bath. Metal ions in the samples were then measured on a single collector ICP-MS platform ELEMENT 2 (Thermo Finnigan, Ringoes, NJ, USA) which allows metals to be measured at concentrations levels as low as 10−12 units, i.e. in the part per trillion (ppt) range.
7
Monensin Treatment Organ Analysis
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At the end of Monensin treatment, implanted mice were perfused with Krebs (- Na+) to eliminate possible contaminations arising from the intravascular Na+ and sacrificed. Their organs were collected, weighed and digest in concentrated HNO3 (70%). Aliquots of the digested samples (0.25 mL), were mineralized by heating under microwave (ETHOS UP Milestone, Bergamo, Italy) following a program which consisted of a ramp to 160 °C in 8 min, followed by 6 min at 160 °C. After mineralization, 4 mL of ultrapure water were added to the sample volumes. The recovered samples were appropriately diluted for ICP-MS (Element-2; Thermo-Finnigan, Rodano (MI), Italy) analysis.
8
Ultrafine Particle Chemical Characterization
9
Quantifying Cellular Gadolinium Content
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Gd content from cell samples was determined using inductively coupled plasma mass spectrometry (ICP-MS) (Element-2; Thermo-Finnigan, Rodano (MI), Italy). Sample digestion was performed using a high-performance Microwave Digestion System (ETHOS UP Milestone, Bergamo, Italy) after the addition of concentrated HNO3 (70%) to cell lysates (1:1), in a final volume of 0.4 mL. The calibration curve was obtained using four Gd absorption standard solutions (Sigma-Aldrich) in the range 0.1–0.004 μg/mL.
10
Boron Quantification in Melanoma Cells
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A375 human melanoma cells were seeded in 6 cm diameter dishes. After 24 h, cells were incubated for 24 h with increasing concentrations of RuCB1 and RuCB2. At the end of the incubation, cells were washed three times with PBS and detached with trypsin/EDTA. A375 cells were resuspended in 200 μL of PBS, sonicated for 30′′ at 30% power in ice and their protein concentration was measured by the Bradford method. Boron amount [μg g−1] in each cell sample was evaluated by Inductively Coupled Mass Spectrometry (ICP-MS) (Element-2; Thermo-Finnigan, Rodano (MI), Italy) at medium mass resolution. Sample digestion was performed with 1 mL of concentrated HNO3 (70%) using a high-performance Microwave Digestion System (ETHOS UP Milestone, Bergamo, Italy). A natural abundance B standard solution was analyzed during sample runs in order to check changing in the systematic bias. The calibration curve was obtained using four B absorption standard solutions (Sigma-Aldrich) in the range 0.2–0.01 μg mL−1.
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